Dihydro-benzo-oxazine and dihydro-pyrido-oxazine derivatives

ABSTRACT

The invention relates to dihydro-benzo-oxazine and dihydro-pyrido-oxazine compounds of the formula (I) and/or pharmaceutically acceptable salts and/or solvates thereof, 
                         
wherein Y, V, W, U, Q, R 1 , R 5 , R 7  and R 30  are as defined in the description. Such compounds are suitable for the treatment of a disorder or disease which is mediated by the activity of the PI3K enzymes.

This application claims priority to U.S. Provisional Application No.61/579,231 filed Dec. 22, 2011, the contents of which are incorporatedherein by reference in their entirety.

FIELD OF THE INVENTION

The invention relates to the preparation and use of newdihydro-benzo-oxazine and dihydro-pyrido-oxazine derivatives as drugcandidates in free form or in pharmaceutically acceptable salt form withvaluable druglike properties, such as e.g. metabolic stability andsuitable pharmacokinetics, form for the modulation, notably theinhibition of the activity or function of the phosphoinositide 3′ OHkinase family (hereinafter PI3K).

BACKGROUND OF THE INVENTION

Members of the phosphoinositide-3 kinase (PI3K) family are involved incell growth, differentiation, survival, cytoskeletal remodeling and thetrafficking of intracellular organelles in many different types of cells(Okkenhaug and Wymann, Nature Rev. Immunol. 3:317 (2003).

To date, eight mammalian PI3Ks have been identified, divided into threemain classes (I, II and III) on the basis of their genetic sequence,structure, adapter molecules, expression, mode of activation, andpreferred substrate.

PI3Kδ is a lipid kinase belonging to the class I PI3K family (PI3K α, β,γ and δ) that generates second messenger signals downstream of tyrosinekinase-linked receptors. PI3Kδ is a heterodimer composed of an adaptorprotein and a p110δ catalytic subunit which convertsphosphatidylinositol-4,5-bis-phosphate (PtdlnsP2) tophosphatidylinositol-3,4,5-tri-phosphate (PtdlnsP3). Effector proteinsinteract with PtdlnsP3 and trigger specific signaling pathways involvedin cell activation, differentiation, migration, and cell survival.

Expression of the p110δ and p110γ catalytic subunits is preferential toleukocytes. Expression is also observed in smooth muscle cells, myocytesand endothelial cells. In contrast, p110α and p110β are expressed by allcell types (Marone et al. Biochimica et Biophysica Acta 1784:159(2008)).

PI3Kδ is associated with B cell development and function (Okkenhaug etal. Science 297:1031 (2002)).

B cells play also a critical role in the pathogenesis of a number ofautoimmune and allergic diseases as well as in the process of transplantrejection (Martin and Chan, Annu. Rev. Immunol. 24:467 (2006)).

Chemotaxis is involved in many autoimmune or inflammatory diseases, inangiogenesis, invasion/metastasis, neurodegeneration or wound healing(Gerard et al. Nat. Immunol. 2:108 (2001)). Temporarily distinct eventsin leukocyte migration in response to chemokines are fully dependent onPI3Kδ and PI3Kγ (Liu et al. Blood 110:1191 (2007)).

PI3Kα and PI3Kβ play an essential role in maintaining homeostasis andpharmacological inhibition of these molecular targets has beenassociated with cancer therapy (Maira et al. Expert Opin. Ther. Targets12:223 (2008)).

PI3Kα is involved in insulin signaling and cellular growth pathways(Foukas et al. Nature 441:366 (2006)). PI3Kδ isoform-selectiveinhibition is expected to avoid potential side effects such ashyperglycemia, and metabolic or growth disregulation.

Parasitic infections still represent one of the most important causes ofmorbidity and mortality worldwide. Among the parasites that cause humanand animal pathology the phylum apicomplexa comprises a group ofvector-borne parasites that is responsible for a wide variety of seriousillnesses including but not limited to malaria, leishmaniasis andtrypanosomiasis. Malaria alone infects 5-10% of humanity and causesaround two million deaths per year. [Schofield et al, “Immunologicalprocesses in malaria pathogenesis”, Nat Rev Imm 2005], [Schofield L,“Intravascular infiltrates and organ-specific inflammation in malariapathogenesis], [Mishra et al, “TLRs in CNS Parasitic infections”, CurrTop Micro Imm 2009], [Bottieau et al, “Therapy of vector-borne protozoaninfections in nonendemic settings”, Expert Rev. Anti infect. Ther.,2011].

Toll-like receptors (TLRs) are germ-line encoded, phylogeneticallyancient molecules that recognize evolutionary conserved structuralrelevant molecules (known as pathogen—associated molecular patterns(PAMPs)) within microbial pathogens. Various different cell typesincluding cells of the immune system express TLRs and are thereby ableto detect the presence of PAMPs. So far 10 functional TLR family members(TLR1-10) have been described in humans, all of which recognize specificPAMP molecules. Following recognition of these specific PAMPs TLRsinduce and orchestrate the immuneresponse of the host to infections withbacteria, viruses, fungi and parasites. [Hedayat et al, “Targeting ofTLRs: a decade of progress in combating infectious disease”, review,Lancet Infectious disease 2011], [Kwai et al, “TLRs and their crosstalkwith other innate receptors in infection and immunity”, review, ImmunityMay-2011].

The immune system of the infected host responds to infection with theTLR induced production of pro-inflammatory cytokines mainly of theT-helper 1 type (Th1). While adequate amounts of these cytokines arebeneficial and required to clear the infection an overproduction ofthese mediators is harmful to the host and associated with immunemediated pathology including neuropathology and tissue damage withsevere and often fatal consequences. One prominent and highly relevantexample of such immune mediated pathology is acute and cerebral malaria(CM) which causes severe clinical symptoms and is often fatal.[Schofield et al, “Immunological processes in malaria pathogenesis”, NatRev Imm 2005], [Schofield L, “Intravascular infiltrates andorgan-specific inflammation in malaria pathogenesis], [Mishra et al,“TLRs in CNS Parasitic infections”, Curr Top Micro Imm 2009], [Bottieauet al, “Therapy of vector-borne protozoan infections in nonendemicsettings”, Expert Rev. Anti infect. Ther., 2011][Hedayat et al,“Targeting of TLRs: a decade of progress in combating infectiousdisease”, review, Lancet Infectious disease 2011]. Despite progress madein treatment and eradication of malaria, the mortality rate that isassociated with severe malaria, including CM remains unacceptably high.Strategies directed solely at the eradication of the parasite in thehost might therefore not be sufficient to prevent neurologicalcomplications and death in all cases of CM. Development of newinnovative adjunct therapeutic strategies to efficiently reduce theCM-associated mortality and morbidity that is caused, in part, byhost-mediated immunopathology remains therefore an urgent medical need.[Higgins et al, “Immunopathogenesis of falciparum malaria: implicationsfor adjunctive therapy in the management of severe and cerebralmalaria”, Expert Rev. Anti Infect. Ther. 2011]

Recently further evidence has been provided that TLR9 plays a key rolein the recognition and response to parasites including but not limitedto Plasmodium, Leishmania, Trypanosoma and Toxoplasma [Gowda et al, “TheNucleosome is the TLR9-specific Immunostimulatory component ofplasmodium falciparum that activates DCs”, PLoS ONE, June 2011],[Peixoto-Rangel et al, “Candidate gene analysis of ocular toxoplasmosisin Brazil: evidence for a role for TLR9”, Mem Inst Oswaldo Cruz 2009],[Pellegrini et al, “The role of TLRs and adoptive immunity in thedevelopment of protective or pathological immune response triggered bythe Trypanosoma cruzi protozoan”, Future Microbiol 2011] and thatinterference with the activation of TLRs including TLR9 represents apromising strategy to prevent the deleterious inflammatory responses insevere and cerebral malaria [Franklin et al, “Therapeutical targeting ofnucleic acid-sensing TLRs prevents experimental cerebral malaria”, PNAS2011]

Malaria is an infectious disease caused by four protozoan parasites:Plasmodium falciparum; Plasmodium vivax; Plasmodium ovale; andPlasmodium malaria. These four parasites are typically transmitted bythe bite of an infected female Anopheles mosquito. Malaria is a problemin many parts of the world and over the last few decades the malariaburden has steadily increased. An estimated 1-3 million people die everyyear from malaria—mostly children under the age of 5. This increase inmalaria mortality is due in part to the fact that Plasmodium falciparum,the deadliest malaria parasite, has acquired resistance against nearlyall available antimalarial drugs, with the exception of the artemisininderivatives.

Leishmaniasis is caused by one or more than 20 varieties of parasiticprotozoa that belong to the genus Leishmania, and is transmitted by thebite of female sand flies. Leishmaniasis is endemic in about 88countries, including many tropical and sub-tropical areas. There arefour main forms of Leishmaniasis. Visceral leishmaniasis, also calledkala-azar, is the most serious form and is caused by the parasiteLeishmania donovani. Patients who develop visceral leishmaniasis can diewithin months unless they receive treatment. The two main therapies forvisceral leishmaniasis are the antimony derivatives sodiumstibogluconate (Pentostam®) and meglumine antimoniate (Glucantim®).Sodium stibogluconate has been used for about 70 years and resistance tothis drug is a growing problem. In addition, the treatment is relativelylong and painful, and can cause undesirable side effects.

Human African Trypanosomiasis, also known as sleeping sickness, is avector-borne parasitic disease. The parasites concerned are protozoabelonging to the Trypanosoma Genus. They are transmitted to humans bytsetse fly (Glossina Genus) bites which have acquired their infectionfrom human beings or from animals harboring the human pathogenicparasites.

Chagas disease (also called American Trypanosomiasis) is another humanparasitic disease that is endemic amongst poor populations on theAmerican continent. The disease is caused by the protozoan parasiteTrypanosoma cruzi, which is transmitted to humans by blood-suckinginsects. The human disease occurs in two stages: the acute stage, whichoccurs shortly after infection and the chronic stage, which can developover many years. Chronic infections result in various neurologicaldisorders, including dementia, damage to the heart muscle and sometimesdilation of the digestive tract, as well as weight loss. Untreated, thechronic disease is often fatal. The drugs currently available fortreating Chagas disease are Nifurtimox and benznidazole. However,problems with these current therapies include their diverse sideeffects, the length of treatment, and the requirement for medicalsupervision during treatment. Furthermore, treatment is really onlyeffective when given during the acute stage of the disease. Resistanceto the two frontline drugs has already occurred. The antifungal agentAmphotericin b has been proposed as a second-line drug, but this drug iscostly and relatively toxic.

Toxoplasmosis is endemic through most of the world, which can infect alarge proportion of the adult population.1,2 However, its prevalencediffers in different countries.3 It is estimated to infect at least 10%of adults in northern temperate countries and more than half of adultsin Mediterranean and tropical contries.4 Toxoplasma gondii is aubiquitous, obligate intracellular protozoan and is considered to be themost common cause of infective retinitis in humans, which depends on avariety of factors, including climate, hygiene, and dietary habits.5-7The course of disease in immunocompetent adults is usually asymptomaticand self-limiting. As soon as infection has occurred, the parasite formslatent cysts in the retina and in other organs of the body, which canreactivate years after the initial infection giving rise to acuteretinochoroiditis and the formation of new retinochoroidal lesions.[Arevalo et al, “Ocular Toxoplasmosis in the developing world”,Internat. Ophthal. Clin 2010]

Neurocysticercosis is the most common parasitic disease of the CNS(incidence ˜2.5 million worldwide) caused by the larvae of Taeniasolium. The disease has a long asymptomatic phase in humanscharacterized by the absence of a detectable inflammatory responsesurrounding the parasite. The overall immune response during theasymptomatic phase is of the Th2 phenotype. However, the destruction oflarvae by therapeutic treatment or by normal parasite attrition causes astrong inflammatory response, often consisting of a chronicgranulomatous reaction and manifestation of typical symptoms of thedisease. The immune response in the CNS of symptomatic patients consistsof an overt Th1 phenotype or a mixed Th1, Th2, and Th3 response,depending upon the absence or presence of granulomas. Thehyperinflammatory response prevailing during the symptomatic phase inthe CNS is responsible for the severe neuropathology and mortalityassociated with neurocysticercosis. [Mishra et al, “TLRs in CNSParasitic infections”, Curr Top Micro Imm 2009]

There is a need to provide new PI3K inhibitors that are good drugcandidates. In particular, compounds of the invention should bindpotently to PI3K whilst showing little affinity for other receptors andshow functional activity as inhibitors. They should be well absorbedfrom the gastrointestinal tract, be metabolically stable and possessfavourable pharmacokinetic properties. When targeted against receptorsin the central nervous system they should cross the blood brain barrierfreely and when targeted selectively against receptors in the peripheralnervous system they should not cross the blood brain barrier. Theyshould be non-toxic and demonstrate few side-effects. Furthermore, theideal drug candidate will exist in a physical form that is stable,non-hygroscopic and easily formulated.

The compounds of the invention show a certain level of selectivityagainst the different paralogs PI3Kα, β, γ and δ. In particular, show acertain level of selectivity for the isoform PI3Kδ.

The compounds of the present invention are therefore potentially usefulin the treatment of a wide range of disorders, particularly disordersincluding but not limited to autoimmune disorders, inflammatorydiseases, allergic diseases, disease or infection associatedimmunopathologies, airway diseases, such as asthma and COPD, transplantrejection, cancers eg of hematopoietic origin or solid tumors.

The invention also relates to the treatment, either alone or incombination, with one or more other pharmacologically active compounds,includes methods of treating conditions, diseases or disorders in whichone or more of the functions of B cells such as antibody production,antigen presentation, cytokine production or lymphoid organogenesis areabnormal or are undesirable including rheumatoid arthritis, pemphigusvulgaris and related diseases, idiopathic thrombocytopenia purpura,systemic lupus erythematosus, multiple sclerosis, myasthenia gravis,Sjögren's syndrome, autoimmune hemolytic anemia, ANCA-associatedvasculitides, cryoglobulinemia, thrombotic thrombocytopenic purpura,chronic autoimmune urticaria, allergy (atopic dermatitis, contactdermatitis, allergic rhinitis), goodpasture's syndrome, AMR(antibody-mediated transplant rejection), B cell-mediated hyperacute,acute and chronic transplant rejection and cancers of haematopoieticorigin including but not limited to multiple myeloma; acute myelogenousleukemia; chronic myelogenous leukemia; lymphocytic leukemia; myeloidleukemia; non-Hodgkin lymphoma; lymphomas; polycythemia vera; essentialthrombocythemia; myelofibrosis with myeloid metaplasia; and Waldenstroem disease as well as in disease or infection associatedimmunopathology.

SUMMARY OF THE INVENTION

The invention relates to dihydro-benzo-oxazine anddihydro-pyrido-oxazine compounds of the formula (I) and/orpharmaceutically acceptable salts and/or solvates thereof,

wherein

-   Y is selected from O or NH;-   V is selected from CR⁵ or N;-   W is selected from CH₂, or O;-   U is selected from N or CH;-   Q is selected from N or CR₆;-   wherein U and Q are not both N;-   R¹ is selected from phenyl, pyridyl, pyrimininyl, pyrazinyl,    pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl,    or    -   —X—R⁴        -   wherein X is selected from C(O), S(O)₂ or CH₂        -   and        -   R⁴ is selected from C₁-C₈-alkyl, halo-C₁-C₈-alkyl,            hydroxy-C₁-C₈-alkyl, C₁-C₈-alkoxy-C₁-C₈-alkyl,            cyano-C₁-C₈-alkyl, N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl,            C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl, phenyl, heterocyclyl,            heterocyclyl-oxy, heterocyclyl-C₁-C₈-alkyl,            C₃-C₁₂-cycloalkyl, C₃-C₁₂-cycloalkyl-oxy,            C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl, heteroaryl, heteroaryl-oxy,            heteroaryl-C₁-C₈-alkyl, hydroxy, C₁-C₈-alkoxy, amino,            N—C₁-C₈-alkyl-amino or N,N-di-C₁-C₈-alkyl-amino,        -   wherein C₁-C₈-alkyl in N—C₁-C₈-alkyl-amino and in            N,N-di-C₁-C₈-alkyl-amino may be unsubstituted or substituted            by halogen, hydroxy or C₁-C₄-alkoxy,        -   wherein C₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyl and in            C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or            substituted by 1-5 substituents selected from halogen,            hydroxy or C₁-C₄-alkoxy;        -   wherein ‘heterocyclyl’ is a 3 to 7 membered saturated or            partially unsaturated monocyclic ring system containing 1 to            3 heteroatoms selected from N, O or S, each of which is            unsubstituted or substituted by 1-5 substituents selected            from oxo, halogen, C₁-C₈-alkyl, halo-C₁-C₈-alkyl,            hydroxy-C₁-C₈-alkyl, hydroxyl, C₁-C₈-alkoxy,            C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,            N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,            halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl or            C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’            can be attached at a heteroatom or a carbon atom and where            the N and/or S heteroatoms can also optionally be oxidized            to various oxidation states,        -   wherein ‘heteroaryl’ is a 3 to 7 membered fully unsaturated            monocyclic ring system containing 1 to 3 heteroatoms            selected from N, O or S, or pyrazolo[1,5-a]pyrimidine or            imidazo[2,1-b]thiazole, each of which is unsubstituted or            substituted by 1-5 substituents selected from halogen,            C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl,            hydroxyl, C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,            N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,            C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,            hydroxy-C₁-C₈-alkyl-carbonyl or            C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heteroaryl’ can            be attached at a heteroatom or a carbon atom and where the N            and/or S heteroatoms can also optionally be oxidized to            various oxidation states;

R⁶ is selected from hydrogen, halogen, C₁-C₄-alkyl, halo-C₁-C₄-alkyl,C₁-C₄-alkoxy, C₁-C₄-alkyl-sulfonyl, C₁-C₄-alkyl-sulfinyl,C₁-C₄-alkyl-sulfanyl, halo-C₁-C₄-alkoxy, C₁-C₄-alkoxy-C₁-C₄-alkyl,amino, N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino;

R⁷ is selected from hydrogen, halogen, cyano, nitro, C₁-C₄-alkyl,halo-C₁-C₄-alkyl, C₁-C₄-alkoxy, N(R⁸)₂-sulfonyl, C₁-C₄-alkyl-sulfonyl,C₁-C₄-alkyl-sulfonyl-amino, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,N—C₁-C₈-alkyl-amino, or N,N-di-C₁-C₈-alkyl-amino;

or R⁶ and R⁷, together are CH═CH—CH═CH,

-   -   wherein R⁸ is independently selected from hydrogen, C₁-C₄-alkyl,        C₁-C₄-alkoxy or two R⁸ together with the nitrogen they are        attached to form a 4 to 7 membered heterocyclic ring containing        1-2 heteroatoms selected from N, O, S, which is unsubstituted or        substituted by 1-3 substituents selected from C₁-C₄-alkyl;

R⁵ is independently selected from H, D, F or C₁-C₂-alkyl;

R³⁰ is independently selected from H, D or F.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is the X-ray Powder Diffraction Pattern of Example F1,crystalline anhydrous form

FIG. 2 is the differential scanning calorimetry graph of Example F1,crystalline anhydrous form

DETAILED DESCRIPTION OF THE INVENTION

Unless specified otherwise, the term “compounds of the presentinvention” refers to compounds of formula (I) and subformulae thereof,salts of the compound, hydrates or solvates of the compounds and/orsalts, as well as all stereoisomers (including diastereoisomers andenantiomers), tautomers and isotopically labeled compounds (includingdeuterium substitutions). Compounds of the present invention furthercomprise polymorphs of compounds of formula (I) (or subformulae thereof)and salts thereof. Where compounds of formula (I) are mentioned, this ismeant to include also the tautomers and N-oxides of the compounds offormula (I).

The invention may be more fully appreciated by reference to thefollowing description, including the following glossary of terms and theconcluding examples. As used herein, the terms “including”, “containing”and “comprising” are used herein in their open, non-limiting sense.

Tautomers, such as tautomers between keto- and enol form, lactam- andlactim form, amid form and imidic acid form or enamine form and imineform, can be present for example in the R¹ portion of compounds offormula (I). Nitrogen containing heterocyclyl and heteroaryl residuesmay form N-oxides.

Where the plural form is used for compounds, salts, and the like, thisis taken to mean also a single compound, salt, or the like.

The general terms used hereinbefore and hereinafter preferably havewithin the context of this disclosure the following meanings, unlessotherwise indicated:

As used herein, the term “alkyl” refers to a fully saturated branched,including single or multiple branching, or unbranched hydrocarbon moietyhaving up to 20 carbon atoms. Unless otherwise provided, alkyl refers tohydrocarbon moieties having 1 to 16 carbon atoms, 1 to 10 carbon atoms,1 to 7 carbon atoms, or 1 to 4 carbon atoms.

Representative examples of alkyl include, but are not limited to,methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl,tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl,2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl,n-decyl and the like. Typically, alkyl groups have 1-7, more preferably1-4 carbons.

As used herein, the term “halo-alkyl” refers to an alkyl as definedherein, which is substituted by one or more halo groups as definedherein. The halo-alkyl can be mono-halo-alkyl, di-halo-alkyl orpoly-halo-alkyl including per-halo-alkyl. A mono-halo-alkyl can have oneiodo, bromo, chloro or fluoro within the alkyl group. Di-halo-alky andpoly-halo-alkyl groups can have two or more of the same halo atoms or acombination of different halo groups within the alkyl. Typically thepoly-halo-alkyl contains up to 12, or 10, or 8, or 6, or 4, or 3, or 2halo groups. Non-limiting examples of halo-alkyl include fluoro-methyl,di-fluoro-methyl, tri-fluoro-methyl, chloro-methyl, di-chloro-methyl,tri-chloro-methyl, penta-fluoro-ethyl, hepta-fluoro-propyl,di-fluoro-chloro-methyl, di-chloro-fluoro-methyl, di-fluoro-ethyl,di-fluoro-propyl, di-chloro-ethyl and dichloro-propyl. A per-halo-alkylrefers to an alkyl having all hydrogen atoms replaced with halo atoms.

As used herein, the term “heterocyclyl” or “heterocyclic” refers to a 3to 7 membered monocyclic or 7 to 10 membered saturated or partiallysaturated ring or ring system, which contains at least one heteroatomselected from N, O and S, where the N and S can also optionally beoxidized to various oxidation states. ‘Heterocyclyl’ can be attached ata heteroatom or a carbon atom. ‘Heterocyclyl’ can include fused orbridged rings as well as spirocyclic rings.

In the context of R⁴, examples of heterocycles include oxiranyl,aziridinyl, oxetanyl, thiethanyl, acetitinyl, pyrrolidinyl,tetrahydrofuranyl, tetrahydrothiophenyl, 2,3-dihydrofuranyl,2,5-dihydrofuranyl, 2,3-dihydrothiophenyl, 1-pyrrolinyl, 2-pyrrolinyl,3-pyrrolinyl, tetrahydropyranyl, piperidinyl, tetrahydrothiopyranyl,morpholinyl, thiomorpholinyl, oxathianyl, dioxanyl, piperazinyl,dihydropyranyl, tetrahydropyridinyl, dihydrothiopyranyl, azepanyl,thiepanyl and oxepanyl.

In the context of R⁸, examples of heterocycles include pyrrolinyl,piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl,tetrahydropyridinyl and azepanyl.

As used herein, the term “heteroaryl” or “heteroarylic” refers to a 4-,5-, 6-, or 7-membered monocyclic, 7-, 8-, 9-, 10-, 11-, or 12-memberedbicyclic or 10-, 11-, 12-, 13-, 14- or 15-membered tricyclic unsaturatedring or ring system—carrying the highest possible number of conjugateddouble bonds in the ring(s), which contains at least one heteroatomselected from N, O and S, wherein the N and S can also optionally beoxidized to various oxidation states. ‘Heteroaryl’ can be attached at aheteroatom or a carbon atom. ‘Heteroaryl’ can include fused or bridgedrings as well as spirocyclic rings. Examples of heteroaryl includefuranyl, thiophenyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl,isothiazolyl, oxazolyl, isoxazolyl, 1,2,5-oxadiazolyl,1,2,4-oxadiazolyl, 1,2,3-oxadiazolyl, 1,3,4-oxadiazolyl,1,2,5-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,3-thiadiazolyl,1,3,4-thiadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl,pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, 1,2,3-triazinyl,1,2,4-triazinyl and 1,3,5-triazinyl.

As used herein, the term “cycloalkyl” refers to saturated or partiallyunsaturated monocyclic, bicyclic or tricyclic hydrocarbon groups of 3-12carbon atoms. Unless otherwise provided, cycloalkyl refers to cyclichydrocarbon groups having between 3 and 10 ring carbon atoms or between3 and 7 ring carbon atoms. Exemplary bicyclic hydrocarbon groups includeoctahydroindyl, decahydronaphthyl. Exemplary tricyclic hydrocarbonbicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl,6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl,bicyclo[2.2.2]octy. Exemplary tetracyclic hydrocarbon groups includeadamantyl.

As used herein, the term “oxy” refers to an —O— linking group.

As used herein, the term “carboxy” or “carboxyl” is —COOH.

As used herein, all substituents are written in a way to show the orderof functional groups (groups) they are composed of. The functionalgroups are defined herein above.

Various enumerated embodiments of the invention are described herein. Itwill be recognized that features specified in each embodiment may becombined with other specified features to provide further embodiments ofthe present invention.

In one embodiment, the invention provides a compound of the formula (I)and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (I′)

wherein R¹, R⁵, R⁷, R³⁰, Y, V, W, U and Q are as defined above.

In one embodiment, the invention provides a compound of the formula (I)and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ia)

wherein R¹, R⁵, R⁷, R³⁰, Y, V, U and Q are as defined above.

In one embodiment, the invention provides a compound of the formula (I)and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ia′)

wherein R¹, R⁵, R⁷, R³⁰, Y, V, U and Q are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ib)

wherein R¹, R⁵, R⁷, R³⁰, V, W, U and Q are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ib′)

wherein R¹, R⁵, R⁷, R³⁰, V, W, U and Q are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ic)

wherein R¹, R⁵, R⁷, U and Q are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ic′)

wherein R¹, R⁵, R⁷, U and Q are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Id)

wherein R¹, R⁵, R⁷, U and Q are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Id′)

wherein R¹, R⁵, R⁷, U and Q are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ie)

wherein R¹, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ie′)

wherein R¹, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (If)

wherein R¹, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (If′)

wherein R¹, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ig)

wherein X, R⁴, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ig′)

wherein X, R⁴, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ih)

wherein X, R⁴, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ih′)

wherein X, R⁴, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ii)

wherein R⁴, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ii′)

wherein R⁴, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ij)

wherein R⁴, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ij′)

wherein R⁴, R⁵, R⁶ and R⁷ are as defined above.

In another embodiment, the invention provides a compound of the formulae(I), (I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie),(Ie′), (If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii), (Ii′), (Ij) or (Ij′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein

R⁴ is selected from C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl,C₁-C₈-alkoxy-C₁-C₈-alkyl, cyano-C₁-C₈-alkyl,N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl, C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl,phenyl, heterocyclyl, heterocyclyl-C₁-C₈-alkyl, C₃-C₁₂-cycloalkyl,heteroaryl, heteroaryl-C₁-C₈-alkyl, C₁-C₈-alkoxy, wherein C₁-C₈-alkyl inN—C₁-C₈-alkyl-amino and in N,N-di-C₁-C₈-alkyl-amino may be unsubstitutedor substituted by halogen, hydroxy or C₁-C₄-alkoxy,

-   -   wherein C₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyl and in        C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or        substituted by halogen, hydroxy or C₁-C₄-alkoxy;    -   wherein ‘heterocyclyl’ is a 3 to 7 membered saturated or        partially unsaturated monocyclic ring system containing 1 to 3        heteroatoms selected from N, O or S, which is unsubstituted or        substituted by 1-5 substituents selected from oxo, halogen,        C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,        C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,        N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,        C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,        hydroxy-C₁-C₈-alkyl-carbonyl or        C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’ can be        attached at a heteroatom or a carbon atom and where the N and/or        S heteroatoms can also optionally be oxidized to various        oxidation states,    -   wherein ‘heteroaryl’ is a 3 to 7 membered fully unsaturated        monocyclic ring system containing 1 to 3 heteroatoms selected        from N, O or S, or pyrazolo[1,5-a]pyrimidine or        imidazo[2,1-b]thiazole, each of which is unsubstituted or        substituted by 1-5 substituents selected from halogen,        C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,        C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,        N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,        C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,        hydroxy-C₁-C₈-alkyl-carbonyl or        C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heteroaryl’ can be        attached at a heteroatom or a carbon atom and where the N and/or        S heteroatoms can also optionally be oxidized to various        oxidation states.

In another embodiment, the invention provides a compound of the formulae(I), (I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie),(Ie′), (If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii), (Ii′), (Ij) or (Ij′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein

R⁴ is selected from C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl,C₁-C₈-alkoxy-C₁-C₈-alkyl, cyano-C₁-C₅-alkyl,N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl, C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl,phenyl, heterocyclyl, heterocyclyl-C₁-C₈-alkyl, C₃-C₁₂-cycloalkyl,heteroaryl, heteroaryl-C₁-C₈-alkyl, C₁-C₈-alkoxy, wherein C₁-C₈-alkyl inN—C₁-C₈-alkyl-amino and in N,N-di-C₁-C₈-alkyl-amino may be unsubstitutedor substituted by halogen, hydroxy or C₁-C₄-alkoxy,

-   -   wherein C₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyl and in        C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or        substituted by halogen, hydroxy or C₁-C₄-alkoxy;    -   wherein ‘heterocyclyl’ is selected from pyrrolidinyl,        tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl,        piperidinyl, tetrahydrothiopyranyl, morpholinyl, dioxanyl or        dihydropyranyl, each of which is unsubstituted or substituted by        1-3 substituents selected from oxo, C₁-C₈-alkyl or        C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’ can be attached at        a heteroatom or a carbon atom and where the N and/or S        heteroatoms can also optionally be oxidized to various oxidation        states, wherein ‘heteroaryl’ is selected from imidazolyl,        pyrazolyl, thiazolyl, oxazolyl, 1,2,5-oxadiazolyl,        1,3,4-oxadiazolyl, pyridyl, pyrimidinyl, pyrazinyl,        pyrazolo[1,5-a]pyrimidine or imidazo[2,1-b]thiazole, each of        which is unsubstituted or substituted by 1-3 substituents        selected from C₁-C₈-alkyl, hydroxyl or amino; wherein        ‘heteroaryl’ can be attached at a heteroatom or a carbon atom        and where the N and/or S heteroatoms can also optionally be        oxidized to various oxidation states.

In another embodiment, the invention provides a compound of the formulae(I), (I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie),(Ie′), (If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii), (Ii′), (Ij) or (Ij′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein R⁶ is selected from halogen, C₁-C₄-alkoxy, C₁-C₄-alkyl-sulfonylor halo-C₁-C₄-alkoxy.

In another embodiment, the invention provides a compound of the formulae(I), (I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie),(Ie′), (If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii), (Ii′), (Ij) or (Ij′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein R⁷ is selected from hydrogen, halogen, cyano, C₁-C₄-alkyl,halo-C₁-C₄-alkyl or C₁-C₄-alkoxy.

In another embodiment, the invention provides a compound of the formulae(I), (I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie),(Ie′), (If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii), (Ii′), (Ij) or (Ij′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein

R⁴ is selected from C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl,C₁-C₈-alkoxy-C₁-C₈-alkyl, cyano-C₁-C₈-alkyl,N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl, C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl,phenyl, heterocyclyl, heterocyclyl-C₁-C₈-alkyl, C₃-C₁₂-cycloalkyl,heteroaryl, heteroaryl-C₁-C₈-alkyl, C₁-C₈-alkoxy, wherein C₁-C₈-alkyl inN—C₁-C₈-alkyl-amino and in N,N-di-C₁-C₈-alkyl-amino may be unsubstitutedor substituted by halogen, hydroxy or C₁-C₄-alkoxy,

-   -   wherein C₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyl and in        C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or        substituted by halogen, hydroxy or C₁-C₄-alkoxy;    -   wherein ‘heterocyclyl’ is a 3 to 7 membered saturated or        partially unsaturated monocyclic ring system containing 1 to 3        heteroatoms selected from N, O or S, which is unsubstituted or        substituted by 1-5 substituents selected from oxo, halogen,        C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,        C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,        N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,        C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,        hydroxy-C₁-C₈-alkyl-carbonyl or        C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’ can be        attached at a heteroatom or a carbon atom and where the N and/or        S heteroatoms can also optionally be oxidized to various        oxidation states,    -   wherein ‘heteroaryl’ is a 3 to 7 membered fully unsaturated        monocyclic ring system containing 1 to 3 heteroatoms selected        from N, O or S, or pyrazolo[1,5-a]pyrimidine or        imidazo[2,1-b]thiazole, each of which is unsubstituted or        substituted by 1-5 substituents selected from halogen,        C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,        C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,        N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,        C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,        hydroxy-C₁-C₈-alkyl-carbonyl or        C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heteroaryl’ can be        attached at a heteroatom or a carbon atom and where the N and/or        S heteroatoms can also optionally be oxidized to various        oxidation states;

and R⁶ is selected from halogen, C₁-C₄-alkoxy, C₁-C₄-alkyl-sulfonyl orhalo-C₁-C₄-alkoxy.

In another embodiment, the invention provides a compound of the formulae(I), (I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie),(Ie′), (If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii), (Ii′), (Ij) or (Ij′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein

R⁴ is selected from C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl,C₁-C₈-alkoxy-C₁-C₈-alkyl, cyano-C₁-C₈-alkyl,N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl, C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl,phenyl, heterocyclyl, heterocyclyl-C₁-C₈-alkyl, C₃-C₁₂-cycloalkyl,heteroaryl, heteroaryl-C₁-C₈-alkyl, C₁-C₈-alkoxy, wherein C₁-C₈-alkyl inN—C₁-C₈-alkyl-amino and in N,N-di-C₁-C₈-alkyl-amino may be unsubstitutedor substituted by halogen, hydroxy or C₁-C₄-alkoxy,

-   -   wherein C₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyl and in        C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or        substituted by halogen, hydroxy or C₁-C₄-alkoxy;    -   wherein ‘heterocyclyl’ is a 3 to 7 membered saturated or        partially unsaturated monocyclic ring system containing 1 to 3        heteroatoms selected from N, O or S, which is unsubstituted or        substituted by 1-5 substituents selected from oxo, halogen,        C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,        C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,        N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,        C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,        hydroxy-C₁-C₈-alkyl-carbonyl or        C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’ can be        attached at a heteroatom or a carbon atom and where the N and/or        S heteroatoms can also optionally be oxidized to various        oxidation states,    -   wherein ‘heteroaryl’ is a 3 to 7 membered fully unsaturated        monocyclic ring system containing 1 to 3 heteroatoms selected        from N, O or S, or pyrazolo[1,5-a]pyrimidine or        imidazo[2,1-b]thiazole, each of which is unsubstituted or        substituted by 1-5 substituents selected from halogen,        C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,        C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,        N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,        C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,        hydroxy-C₁-C₈-alkyl-carbonyl or        C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heteroaryl’ can be        attached at a heteroatom or a carbon atom and where the N and/or        S heteroatoms can also optionally be oxidized to various        oxidation states;

and R⁷ is selected from hydrogen, halogen, cyano, C₁-C₄-alkyl,halo-C₁-C₄-alkyl or C₁-C₄-alkoxy.

In another embodiment, the invention provides a compound of the formulae(I), (I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie),(Ie′), (If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii), (Ii′), (Ij) or (Ij′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein

R⁴ is selected from C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl,C₁-C₈-alkoxy-C₁-C₈-alkyl, cyano-C₁-C₈-alkyl,N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl, C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl,phenyl, heterocyclyl, heterocyclyl-C₁-C₈-alkyl, C₃-C₁₂-cycloalkyl,heteroaryl, heteroaryl-C₁-C₈-alkyl, C₁-C₈-alkoxy, wherein C₁-C₈-alkyl inN—C₁-C₈-alkyl-amino and in N,N-di-C₁-C₈-alkyl-amino may be unsubstitutedor substituted by halogen, hydroxy or C₁-C₄-alkoxy,

-   -   wherein C₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyl and in        C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or        substituted by halogen, hydroxy or C₁-C₄-alkoxy;    -   wherein ‘heterocyclyl’ is a 3 to 7 membered saturated or        partially unsaturated monocyclic ring system containing 1 to 3        heteroatoms selected from N, O or S, which is unsubstituted or        substituted by 1-5 substituents selected from oxo, halogen,        C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,        C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,        N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,        C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,        hydroxy-C₁-C₈-alkyl-carbonyl or        C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’ can be        attached at a heteroatom or a carbon atom and where the N and/or        S heteroatoms can also optionally be oxidized to various        oxidation states,    -   wherein ‘heteroaryl’ is a 3 to 7 membered fully unsaturated        monocyclic ring system containing 1 to 3 heteroatoms selected        from N, O or S, or pyrazolo[1,5-a]pyrimidine or        imidazo[2,1-b]thiazole, each of which is unsubstituted or        substituted by 1-5 substituents selected from halogen,        C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,        C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,        N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,        C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,        hydroxy-C₁-C₈-alkyl-carbonyl or        C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heteroaryl’ can be        attached at a heteroatom or a carbon atom and where the N and/or        S heteroatoms can also optionally be oxidized to various        oxidation states;

R⁶ is selected from halogen, C₁-C₄-alkoxy, C₁-C₄-alkyl-sulfonyl orhalo-C₁-C₄-alkoxy and R⁷ is selected from hydrogen, halogen, cyano,C₁-C₄-alkyl, halo-C₁-C₄-alkyl or C₁-C₄-alkoxy.

In another embodiment individual compounds according to the inventionare those listed in the Examples section below.

As used herein, the term “an optical isomer” or “a stereoisomer” refersto any of the various stereo isomeric configurations which may exist fora given compound of the present invention and includes geometricisomers. It is understood that a substituent may be attached at a chiralcenter of a carbon atom. The term “chiral” refers to molecules whichhave the property of non-superimposability on their mirror imagepartner, while the term “achiral” refers to molecules which aresuperimposable on their mirror image partner. Therefore, the inventionincludes enantiomers, diastereomers or racemates of the compound.“Enantiomers” are a pair of stereoisomers that are non-superimposablemirror images of each other. A 1:1 mixture of a pair of enantiomers is a“racemic” mixture. The term is used to designate a racemic mixture whereappropriate. “Diastereoisomers” are stereoisomers that have at least twoasymmetric atoms, but which are not mirror-images of each other. Theabsolute stereochemistry is specified according to theCahn-Ingold-Prelog R-S system. When a compound is a pure enantiomer thestereochemistry at each chiral carbon may be specified by either R or S.Resolved compounds whose absolute configuration is unknown can bedesignated (+) or (−) depending on the direction (dextro- orlevorotatory) which they rotate plane polarized light at the wavelengthof the sodium D line. Certain compounds described herein contain one ormore asymmetric centers or axes and may thus give rise to enantiomers,diastereomers, and other stereoisomeric forms that may be defined, interms of absolute stereochemistry, as (R)- or (S)-.

Depending on the choice of the starting materials and procedures, thecompounds can be present in the form of one of the possible isomers oras mixtures thereof, for example as pure optical isomers, or as isomermixtures, such as racemates and diastereoisomer mixtures, depending onthe number of asymmetric carbon atoms. The present invention is meant toinclude all such possible isomers, including racemic mixtures,diasteriomeric mixtures and optically pure forms. Optically active (R)-and (S)-isomers may be prepared using chiral synthons or chiralreagents, or resolved using conventional techniques. If the compoundcontains a double bond, the substituent may be E or Z configuration. Ifthe compound contains a disubstituted cycloalkyl, the cycloalkylsubstituent may have a cis- or trans-configuration. All tautomeric formsare also intended to be included.

As used herein, the terms “salt” or “salts” refers to an acid additionor base addition salt of a compound of the invention. “Salts” include inparticular “pharmaceutical acceptable salts”. The term “pharmaceuticallyacceptable salts” refers to salts that retain the biologicaleffectiveness and properties of the compounds of this invention and,which typically are not biologically or otherwise undesirable. In manycases, the compounds of the present invention are capable of formingacid and/or base salts by virtue of the presence of amino and/orcarboxyl groups or groups similar thereto.

Pharmaceutically acceptable acid addition salts can be formed withinorganic acids and organic acids, e.g., acetate, aspartate, benzoate,besylate, bromide/hydrobromide, bicarbonate/carbonate,bisulfate/sulfate, camphorsulfonate, chloride/hydrochloride,chlortheophyllonate, citrate, ethandisulfonate, fumarate, gluceptate,gluconate, glucuronate, hippurate, hydroiodide/iodide, isethionate,lactate, lactobionate, laurylsulfate, malate, maleate, malonate,mandelate, mesylate, methylsulphate, naphthoate, napsylate, nicotinate,nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, polygalacturonate,propionate, stearate, succinate, sulfosalicylate, tartrate, tosylate andtrifluoroacetate salts.

Inorganic acids from which salts can be derived include, for example,hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like. Organic acids from which salts can bederived include, for example, acetic acid, propionic acid, glycolicacid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaricacid, tartaric acid, citric acid, benzoic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid,sulfosalicylic acid, and the like. Pharmaceutically acceptable baseaddition salts can be formed with inorganic and organic bases.

Inorganic bases from which salts can be derived include, for example,ammonium salts and metals from columns I to XII of the periodic table.In certain embodiments, the salts are derived from sodium, potassium,ammonium, calcium, magnesium, iron, silver, zinc, and copper;particularly suitable salts include ammonium, potassium, sodium, calciumand magnesium salts.

Organic bases from which salts can be derived include, for example,primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines, basic ionexchange resins, and the like. Certain organic amines includeisopropylamine, benzathine, cholinate, diethanolamine, diethylamine,lysine, meglumine, piperazine and tromethamine.

The pharmaceutically acceptable salts of the present invention can besynthesized from a basic or acidic moiety, by conventional chemicalmethods. Generally, such salts can be prepared by reacting free acidforms of these compounds with a stoichiometric amount of the appropriatebase (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate or thelike), or by reacting free base forms of these compounds with astoichiometric amount of the appropriate acid. Such reactions aretypically carried out in water or in an organic solvent, or in a mixtureof the two. Generally, use of non-aqueous media like ether, ethylacetate, ethanol, isopropanol, or acetonitrile is desirable, wherepracticable. Lists of additional suitable salts can be found, e.g., in“Remington's Pharmaceutical Sciences”, 20th ed., Mack PublishingCompany, Easton, Pa., (1985); and in “Handbook of Pharmaceutical Salts:Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH,Weinheim, Germany, 2002).

Any formula given herein is also intended to represent unlabeled formsas well as isotopically labeled forms of the compounds. Isotopicallylabeled compounds have structures depicted by the formulas given hereinexcept that one or more atoms are replaced by an atom having a selectedatomic mass or mass number. Examples of isotopes that can beincorporated into compounds of the invention include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine,such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸F, ³¹P, ³²P, ³⁵S, ³⁶Cl, ¹²⁵Irespectively. The invention includes various isotopically labeledcompounds as defined herein, for example those into which radioactiveisotopes, such as ³H and ¹⁴C, or those into which non-radioactiveisotopes, such as ²H and ¹³C are present. Such isotopically labelledcompounds are useful in metabolic studies (with ¹⁴C), reaction kineticstudies (with, for example ²H or ³H), detection or imaging techniques,such as positron emission tomography (PET) or single-photon emissioncomputed tomography (SPECT) including drug or substrate tissuedistribution assays, or in radioactive treatment of patients. Inparticular, an ¹⁸F or labeled compound may be particularly desirable forPET or SPECT studies. Isotopically-labeled compounds of formula (I) cangenerally be prepared by conventional techniques known to those skilledin the art or by processes analogous to those described in theaccompanying Examples and Preparations using an appropriateisotopically-labeled reagent in place of the non-labeled reagentpreviously employed.

Further, substitution with heavier isotopes, particularly deuterium(i.e., ²H or D) may afford certain therapeutic advantages resulting fromgreater metabolic stability, for example increased in vivo half-life orreduced dosage requirements or an improvement in therapeutic index. Itis understood that deuterium in this context is regarded as asubstituent of a compound of the formula (I). The concentration of sucha heavier isotope, specifically deuterium, may be defined by theisotopic enrichment factor. The term “isotopic enrichment factor” asused herein means the ratio between the isotopic abundance and thenatural abundance of a specified isotope. If a substituent in a compoundof this invention is denoted deuterium, such compound has an isotopicenrichment factor for each designated deuterium atom of at least 3500(52.5% deuterium incorporation at each designated deuterium atom), atleast 4000 (60% deuterium incorporation), at least 4500 (67.5% deuteriumincorporation), at least 5000 (75% deuterium incorporation), at least5500 (82.5% deuterium incorporation), at least 6000 (90% deuteriumincorporation), at least 6333.3 (95% deuterium incorporation), at least6466.7 (97% deuterium incorporation), at least 6600 (99% deuteriumincorporation), or at least 6633.3 (99.5% deuterium incorporation).

Pharmaceutically acceptable solvates in accordance with the inventioninclude those wherein the solvent of crystallization may be isotopicallysubstituted, e.g. D₂O, d₆-acetone, DMSO-d6.

Compounds of the invention, i.e. compounds of formula (I) that containgroups capable of acting as donors and/or acceptors for hydrogen bondsmay be capable of forming co-crystals with suitable co-crystal formers.These co-crystals may be prepared from compounds of formula (I) by knownco-crystal forming procedures. Such procedures include grinding,heating, co-subliming, co-melting, or contacting in solution compoundsof formula (I) with the co-crystal former under crystallizationconditions and isolating co-crystals thereby formed. Suitable co-crystalformers include those described in WO 2004/078163. Hence the inventionfurther provides co-crystals comprising a compound of formula (I).

As used herein, the term “pharmaceutically acceptable carrier” includesany and all solvents, dispersion media, coatings, surfactants,antioxidants, preservatives (e.g., antibacterial agents, antifungalagents), isotonic agents, absorption delaying agents, salts,preservatives, drug stabilizers, binders, excipients, disintegrationagents, lubricants, sweetening agents, flavoring agents, dyes, and thelike and combinations thereof, as would be known to those skilled in theart (see, for example, Remington's Pharmaceutical Sciences, 18th Ed.Mack Printing Company, 1990, pp. 1289-1329). Except insofar as anyconventional carrier is incompatible with the active ingredient, its usein the therapeutic or pharmaceutical compositions is contemplated.

The term “a therapeutically effective amount” of a compound of thepresent invention refers to an amount of the compound of the presentinvention that will elicit the biological or medical response of asubject, for example, reduction or inhibition of an enzyme or a proteinactivity, or ameliorate symptoms, alleviate conditions, slow or delaydisease progression, or prevent a disease, etc. In one non-limitingembodiment, the term “a therapeutically effective amount” refers to theamount of the compound of the present invention that, when administeredto a subject, is effective to (1) at least partially alleviate, inhibit,prevent and/or ameliorate a condition, or a disorder or a disease (i)mediated by PI3K or (ii) associated with PI3K activity, or (iii)characterized by activity (normal or abnormal) of PI3K or (2) reduce orinhibit the activity of PI3K or (3) reduce or inhibit the expression ofPI3K. In another non-limiting embodiment, the term “a therapeuticallyeffective amount” refers to the amount of the compound of the presentinvention that, when administered to a cell, or a tissue, or anon-cellular biological material, or a medium, is effective to at leastpartially reducing or inhibiting the activity of PI3K; or at leastpartially reducing or inhibiting the expression of PI3K. The meaning ofthe term “a therapeutically effective amount” as illustrated in theabove embodiment for PI3K also applies by the same means to any otherrelevant proteins/peptides/enzymes.

As used herein, the term “subject” refers to an animal. Typically theanimal is a mammal. A subject also refers to for example, primates(e.g., humans, male or female), cows, sheep, goats, horses, dogs, cats,rabbits, rats, mice, fish, birds and the like. In certain embodiments,the subject is a primate. In yet other embodiments, the subject is ahuman.

As used herein, the term “inhibit”, “inhibition” or “inhibiting” refersto the reduction or suppression of a given condition, symptom, ordisorder, or disease, or a significant decrease in the baseline activityof a biological activity or process.

As used herein, the term “treat”, “treating” or “treatment” of anydisease or disorder refers in one embodiment, to ameliorating thedisease or disorder (i.e., slowing or arresting or reducing thedevelopment of the disease or at least one of the clinical symptomsthereof). In another embodiment “treat”, “treating” or “treatment”refers to alleviating or ameliorating at least one physical parameterincluding those which may not be discernible by the patient. In yetanother embodiment, “treat”, “treating” or “treatment” refers tomodulating the disease or disorder, either physically, (e.g.,stabilization of a discernible symptom), physiologically, (e.g.,stabilization of a physical parameter), or both. In yet anotherembodiment, “treat”, “treating” or “treatment” refers to preventing ordelaying the onset or development or progression of the disease ordisorder.

As used herein, a subject is “in need of” a treatment if such subjectwould benefit biologically, medically or in quality of life from suchtreatment.

As used herein, the term “a,” “an,” “the” and similar terms used in thecontext of the present invention (especially in the context of theclaims) are to be construed to cover both the singular and plural unlessotherwise indicated herein or clearly contradicted by the context.

All methods described herein can be performed in any suitable orderunless otherwise indicated herein or otherwise clearly contradicted bycontext. The use of any and all examples, or exemplary language (e.g.“such as”) provided herein is intended merely to better illuminate theinvention and does not pose a limitation on the scope of the inventionotherwise claimed.

Any asymmetric atom (e.g., carbon or the like) of the compound(s) of thepresent invention can be present in racemic or enantiomericallyenriched, for example the (R)-, (S)- or (R,S)-configuration. In certainembodiments, each asymmetric atom has at least 50% enantiomeric excess,at least 60% enantiomeric excess, at least 70% enantiomeric excess, atleast 80% enantiomeric excess, at least 90% enantiomeric excess, atleast 95% enantiomeric excess, or at least 99% enantiomeric excess inthe (R)- or (S)-configuration. Substituents at atoms with unsaturateddouble bonds may, if possible, be present in cis-(Z)- or trans-(E)-form.

Accordingly, as used herein a compound of the present invention can bein the form of one of the possible isomers, rotamers, atropisomers,tautomers or mixtures thereof, for example, as substantially puregeometric (cis or trans) isomers, diastereomers, optical isomers(antipodes), racemates or mixtures thereof.

Any resulting mixtures of isomers can be separated on the basis of thephysicochemical differences of the constituents, into the pure orsubstantially pure geometric or optical isomers, diastereomers,racemates, for example, by chromatography and/or fractionalcrystallization.

Any resulting racemates of final products or intermediates can beresolved into the optical antipodes by known methods, e.g., byseparation of the diastereomeric salts thereof, obtained with anoptically active acid or base, and liberating the optically activeacidic or basic compound. In particular, a basic moiety may thus beemployed to resolve the compounds of the present invention into theiroptical antipodes, e.g., by fractional crystallization of a salt formedwith an optically active acid, e.g., tartaric acid, dibenzoyl tartaricacid, diacetyl tartaric acid, di-O,O′-p-toluoyl tartaric acid, mandelicacid, malic acid or camphor-10-sulfonic acid. Racemic products can alsobe resolved by chiral chromatography, e.g., high pressure liquidchromatography (HPLC) using a chiral adsorbent.

Furthermore, the compounds of the present invention, including theirsalts, can also be obtained in the form of their hydrates, or includeother solvents used for their crystallization. The compounds of thepresent invention may inherently or by design form solvates withpharmaceutically acceptable solvents (including water); therefore, it isintended that the invention embrace both solvated and unsolvated forms.The term “solvate” refers to a molecular complex of a compound of thepresent invention (including pharmaceutically acceptable salts thereof)with one or more solvent molecules. Such solvent molecules are thosecommonly used in the pharmaceutical art, which are known to be innocuousto the recipient, e.g., water, ethanol, and the like. The term “hydrate”refers to the complex where the solvent molecule is water.

The compounds of the present invention, including salts, hydrates andsolvates thereof, may inherently or by design form polymorphs.

Typically, the compounds of formula (I) can be prepared according to themethods provided infra.

In one embodiment, the invention relates to a process for manufacturinga compound of formula (I) (Method A) comprising steps a, b, c, d.

The compound of formula (I) is obtained via the step c of deprotectingPG¹ from the compound of formula (F), wherein PG¹ represents a suitableprotecting group, such as a Boc group, and the other substituents are asdefined above,

followed by coupling reaction step d withR¹-Act¹,

step c1: Where R¹ is —C(O)—R⁴, or —S(O)₂—R⁴, wherein R⁴ is definedabove, and Act¹ represents an activating group or a hydroxy group: Thecoupling reaction is an amide, urea, carbamic ester or sulfonamidformation. There are many known ways of preparing amides, urea carbamicesters or sulfonamids. The coupling reaction step can be carried outwith Act¹ representing an activating group, preferably in a one stepprocedure or with Act¹ representing a hydroxy group either involving aone or two step procedure. For examples of amide bond formations, seeMantalbetti, C. A. G. N and Falque, V., Amide bond formation and peptidecoupling, Tetrahedron, 2005, 61(46), pp 10827-10852 and references citedtherein. For examples of urea synthesis, see Sartori, G.; Maggi, R.Acyclic and cyclic ureas, Science of Synthesis (2005), 18, 665-758;Gallou, Isabelle. Unsymmetrical ureas Synthetic methodologies andapplication in drug design, Organic Preparations and ProceduresInternational (2007), 39(4), 355-383. For examples of carbamatesynthesis see Adams, Philip; Baron, Frank A. Esters of carbamic acid,Chemical Reviews (1965), 65(5), 567-602. The examples provided hereinare thus not intended to be exhaustive, but merely illustrative;

step c2: Where R¹ is selected from phenyl, pyridyl, pyrimidinyl,pyrazinyl, pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl or1,3,5-triazinyl and Act¹ represents halogen, particularly iodo or bromo:The coupling reaction is carried out in the presence of an amine basesuch as N,N-diisopropylethylamine. The reaction is carried out in thepresence of an organic solvent or without a solvent under microwaveheating. Alternatively, the reaction is carried out under customaryBuchwald-Hartwig conditions such as the conditions described above. Thereaction is preferably carried out under an inert gas such as nitrogenor argon.

The compound of formula (F) is obtained via the step b of coupling thecompound of formula (C), wherein PG¹ represents a suitable protectinggroup, such as a Boc, and the other substituents are as defined above,

with a compound of formula (D), wherein X′ represents halogen, such asiodo or bromo and the other substituents are as defined above,

under customary Buchwald-Hartwig conditions using a suitable Pdcatalyst/ligand combination such asPd₂(dba)₃/2-(dicyclohexylphosphino)biphenyl orPd₂(dba)₃/2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-biphenyl,Pd₂(dba)₃/X-Phos, Pd₂(dba)₃/(rac)-BINAP, Pd(OAc)₂/(rac)-BINAP orbis(tri-t-butylphosphine)palladium and a suitable base, such as NaOtBu,Cs₂CO₃ or K₃PO₄ and organic solvent such as toluene, dioxane or THF. Thereaction is stirred at a temperature of approximately 60-140° C., forexample at 100° C. to 110° C. and is optionally performed in a microwavereactor. The reaction is preferably carried out under an inert gas suchas nitrogen or argon.

The compound of formula (C) is obtained via the step a of coupling thecompound of formula (A), wherein the substituents are as defined abovewith a compound of formula (B), wherein PG¹ represents a suitableprotecting group, such as a Boc group and Act² is an activating group orH, and the other substituents are as defined above,

step a1: Where Y is O and Act² represents an activating group such as amesylate: The reaction takes place in the presence of a suitable basesuch as sodium hydroxide (NaH), K₂CO₃ or potassium t-butoxide (tBuOK) ina suitable polar organic solvent such as DMF, THF,2-methyltetrahydrofuran or Dioxane at a suitable temperature such asrt—100° C.

step a2: Where Y is O and Act² represents H: The reaction takes placeusing customary Mitsunobu conditions, for example using Ph₃P and DEAD inorganic solvent such as THF under inert gas conditions at elevatedtemperature such as 70° C.

step a3: Where Y is NH and Act² represents H: A base promotedphosphonium coupling reaction is employed, whereby a compound of theformula (A) in a suitable solvent such as acetonitrile is reacted with aphosphonium salt such asbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(BOP) in the presence of a base such as1,8-diaza-7-bicyclo[5.4.0]undecene (DBU) followed by addition of acompound of the formula (B). The reaction mixture is stirred at atemperature of 20° C. to 100° C.

In another embodiment, the invention relates to a process formanufacturing a compound of formula (I) (Method A-a) comprising steps aand b as defined above for Method A, using a compound of formula (B)wherein PG¹ represents R¹.

In another embodiment, the invention relates to a process formanufacturing a compound of formula (I) (Method B) comprising steps e,b, f, a, c and d.

The compound of formula (I) is obtained via the steps c and d asdescribed above for Method A from the compound of formula (F).

The compound of formula (F) is obtained via the step f of deprotectingPG² from the compound of formula (E), wherein PG² is a suitableprotecting group, such as a silyl protecting group, and the othersubstituents are as defined above

followed by coupling reaction step a, as described above for Method A,with the compound of formula (B).

The compound of formula (E) is obtained via the step e of protecting thecompound of formula (A) with a suitable protecting group PG², followedby from the compound of formula (E), wherein PG² is a suitableprotecting group, such as a silyl protecting group, followed by couplingreaction step b, as described above for Method A with the compound offormula (D).

In another embodiment, the invention relates to a process formanufacturing a compound of formula (I) (Method B-a) comprising steps e,b and f as defined above for Method B, using a compound of formula (B)wherein PG¹ represents R¹.

In another embodiment, the invention relates to a process formanufacturing a compound of formula (I) (Method C) using a compound offormula (A′), wherein X″ represents halogen and the other substituentsare as defined above

comprising steps b, c and d as defined above for Method B, using acompound of formula (B) and a modified step a4:

step a4: Where Y is NH and Act² is H: The reaction takes place in thepresence of a suitable base such as for example potassium carbonate or asuitable amine base such as triethylamine or N,N-diisopropylethylamineat elevated temperature such as 100° C. to 140° C. Alternatively, thereaction is carried out under customary Buchwald-Hartwig conditions suchas the conditions described above. The reaction is preferably carriedout under an inert gas such as nitrogen or argon.

In another embodiment, the invention relates to a process formanufacturing a compound of formula (I) (Method C-a) comprising steps b,a4, c and d as defined above for Method B¹, using a compound of formula(B) wherein PG¹ represents R¹.

The term “activating group” as used herein relates to a group that canactivate a carboxylic acid, carbonic acid or carbamic acid derivative,for coupling with an amine moiety to form an amide, urea or carbamicester moiety, respectively (Act¹) or to a group that can activate ahydroxy group for coupling with another hydroxy moiety to form an ether(Act²).

Groups that can activate a carboxylic acid, carbonic acid or carbamicacid derivative, for coupling with an amine moiety to form an amide,urea or carbamic ester moiety are chlorides, or groups resulting fromthe reaction of the acid derivative with an activating agent. Suitableactivating agents are known to the skilled person, examples of suchactivating reagents are carbodiimide derivatives, pentafluorophenylester derivatives, triazole derivatives, imidazole derivatives.

Groups that can activate a hydroxy group for coupling with anotherhydroxy moiety to form an ether are groups are known to the skilledperson, examples of such activating groups are mesylates and tosylates.

The term “protecting group” as used herein relates to a group thatprotects a functional group which is present in the starting materialsand is not intended to take part in the reaction. In additional processsteps, carried out as desired, functional groups of the startingcompounds which should not take part in the reaction may be present inunprotected form or may be protected for example by one or moreprotecting groups. The protecting groups are then wholly or partlyremoved according to one of the known methods. Protecting groups, andthe manner in which they are introduced and removed are described, forexample, in “Protective Groups in Organic Chemistry”, Plenum Press,London, New York 1973, and in “Methoden der organischen Chemie”,Houben-Weyl, 4th edition, Vol. 15/1, Georg-Thieme-Verlag, Stuttgart 1974and in Theodora W. Greene, “Protective Groups in Organic Synthesis”,John Wiley & Sons, New York 1981. A characteristic of protecting groupsis that they can be removed readily, i.e. without the occurrence ofundesired secondary reactions, for example by solvolysis, reduction,photolysis or alternatively under physiological conditions.

The invention further includes any variant of the present processes, inwhich an intermediate product obtainable at any stage thereof is used asstarting material and the remaining steps are carried out, or in whichthe starting materials are formed in situ under the reaction conditions,or in which the reaction components are used in the form of their saltsor optically pure material.

Compounds of the invention and intermediates can also be converted intoeach other according to methods generally known to those skilled in theart.

Intermediates and final products can be worked up and/or purifiedaccording to standard methods, e.g. using chromatographic methods,distribution methods, (re-) crystallization, and the like.

The following applies in general to all processes mentioned hereinbefore and hereinafter.

All the above-mentioned process steps can be carried out under reactionconditions that are known to those skilled in the art, including thosementioned specifically, in the absence or, customarily, in the presenceof solvents or diluents, including, for example, solvents or diluentsthat are inert towards the reagents used and dissolve them, in theabsence or presence of catalysts, condensation or neutralizing agents,for example ion exchangers, such as cation exchangers, e.g. in the H+form, depending on the nature of the reaction and/or of the reactants atreduced, normal or elevated temperature, for example in a temperaturerange of from about −100° C. to about 190° C., including, for example,from approximately −80° C. to approximately 150° C., for example at from−80 to −60° C., at room temperature, at from −20 to 40° C. or at refluxtemperature, under atmospheric pressure or in a closed vessel, whereappropriate under pressure, and/or in an inert atmosphere, for exampleunder an argon or nitrogen atmosphere.

At all stages of the reactions, mixtures of isomers that are formed canbe separated into the individual isomers, for example diastereoisomersor enantiomers, or into any desired mixtures of isomers, for exampleracemates or mixtures of diastereoisomers, for example analogously tothe methods described herein above.

The solvents from which those solvents that are suitable for anyparticular reaction may be selected include those mentioned specificallyor, for example, water, esters, such as lower alkyl-lower alkanoates,for example ethyl acetate, ethers, such as aliphatic ethers, for examplediethyl ether, or cyclic ethers, for example tetrahydrofuran or dioxane,liquid aromatic hydrocarbons, such as benzene or toluene, alcohols, suchas methanol, ethanol or 1- or 2-propanol, nitriles, such asacetonitrile, halogenated hydrocarbons, such as methylene chloride orchloroform, acid amides, such as dimethylformamide or dimethylacetamide, bases, such as heterocyclic nitrogen bases, for examplepyridine or N-methylpyrrolidin-2-one, carboxylic acid anhydrides, suchas lower alkanoic acid anhydrides, for example acetic anhydride, cyclic,linear or branched hydrocarbons, such as cyclohexane, hexane orisopentane, methycyclohexane, or mixtures of those solvents, for exampleaqueous solutions, unless otherwise indicated in the description of theprocesses. Such solvent mixtures may also be used in working up, forexample by chromatography or partitioning.

The compounds, including their salts, may also be obtained in the formof hydrates, or their crystals may, for example, include the solventused for crystallization. Different crystalline forms may be present.

The invention relates also to those forms of the process in which acompound obtainable as an intermediate at any stage of the process isused as starting material and the remaining process steps are carriedout, or in which a starting material is formed under the reactionconditions or is used in the form of a derivative, for example in aprotected form or in the form of a salt, or a compound obtainable by theprocess according to the invention is produced under the processconditions and processed further in situ.

In another aspect, the present invention provides a pharmaceuticalcomposition comprising a compound of the present invention, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier. The pharmaceutical composition can be formulated forparticular routes of administration such as oral administration,parenteral administration, and rectal administration, etc. In addition,the pharmaceutical compositions of the present invention can be made upin a solid form (including without limitation capsules, tablets, pills,granules, powders or suppositories), or in a liquid form (includingwithout limitation solutions, suspensions or emulsions). Thepharmaceutical compositions can be subjected to conventionalpharmaceutical operations such as sterilization and/or can containconventional inert diluents, lubricating agents, or buffering agents, aswell as adjuvants, such as preservatives, stabilizers, wetting agents,emulsifers and buffers, etc.

Typically, the pharmaceutical compositions are tablets or gelatincapsules comprising the active ingredient together with

a) diluents, e.g., lactose, dextrose, sucrose, mannitol, sorbitol,cellulose and/or glycine;

b) lubricants, e.g., silica, talcum, stearic acid, its magnesium orcalcium salt and/or polyethyleneglycol; for tablets also

c) binders, e.g., magnesium aluminum silicate, starch paste, gelatin,tragacanth, methylcellulose, sodium carboxymethylcellulose and/orpolyvinylpyrrolidone; if desired

d) disintegrants, e.g., starches, agar, alginic acid or its sodium salt,or effervescent mixtures; and/or

e) absorbents, colorants, flavors and sweeteners.

Tablets may be either film coated or enteric coated according to methodsknown in the art.

Suitable compositions for oral administration include an effectiveamount of a compound of the invention in the form of tablets, lozenges,aqueous or oily suspensions, dispersible powders or granules, emulsion,hard or soft capsules, or syrups or elixirs. Compositions intended fororal use are prepared according to any method known in the art for themanufacture of pharmaceutical compositions and such compositions cancontain one or more agents selected from the group consisting ofsweetening agents, flavoring agents, coloring agents and preservingagents in order to provide pharmaceutically elegant and palatablepreparations. Tablets may contain the active ingredient in admixturewith nontoxic pharmaceutically acceptable excipients which are suitablefor the manufacture of tablets. These excipients are, for example, inertdiluents, such as calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate; granulating and disintegrating agents,for example, corn starch, or alginic acid; binding agents, for example,starch, gelatin or acacia; and lubricating agents, for example magnesiumstearate, stearic acid or talc. The tablets are uncoated or coated byknown techniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate can be employed. Formulations fororal use can be presented as hard gelatin capsules wherein the activeingredient is mixed with an inert solid diluent, for example, calciumcarbonate, calcium phosphate or kaolin, or as soft gelatin capsuleswherein the active ingredient is mixed with water or an oil medium, forexample, peanut oil, liquid paraffin or olive oil.

Certain injectable compositions are aqueous isotonic solutions orsuspensions, and suppositories are advantageously prepared from fattyemulsions or suspensions. Said compositions may be sterilized and/orcontain adjuvants, such as preserving, stabilizing, wetting oremulsifying agents, solution promoters, salts for regulating the osmoticpressure and/or buffers. In addition, they may also contain othertherapeutically valuable substances. Said compositions are preparedaccording to conventional mixing, granulating or coating methods,respectively, and contain about 0.1-75%, or contain about 1-50%, of theactive ingredient.

Suitable compositions for transdermal application include an effectiveamount of a compound of the invention with a suitable carrier. Carrierssuitable for transdermal delivery include absorbable pharmacologicallyacceptable solvents to assist passage through the skin of the host. Forexample, transdermal devices are in the form of a bandage comprising abacking member, a reservoir containing the compound optionally withcarriers, optionally a rate controlling barrier to deliver the compoundof the skin of the host at a controlled and predetermined rate over aprolonged period of time, and means to secure the device to the skin.

Suitable compositions for topical application, e.g., to the skin andeyes, include aqueous solutions, suspensions, ointments, creams, gels orsprayable formulations, e.g., for delivery by aerosol or the like. Suchtopical delivery systems will in particular be appropriate for dermalapplication, e.g., for the treatment of skin cancer, e.g., forprophylactic use in sun creams, lotions, sprays and the like. They arethus particularly suited for use in topical, including cosmetic,formulations well-known in the art. Such may contain solubilizers,stabilizers, tonicity enhancing agents, buffers and preservatives.

As used herein a topical application may also pertain to an inhalationor to an intranasal application. They may be conveniently delivered inthe form of a dry powder (either alone, as a mixture, for example a dryblend with lactose, or a mixed component particle, for example withphospholipids) from a dry powder inhaler or an aerosol spraypresentation from a pressurised container, pump, spray, atomizer ornebuliser, with or without the use of a suitable propellant.

The present invention further provides anhydrous pharmaceuticalcompositions and dosage forms comprising the compounds of the presentinvention as active ingredients, since water may facilitate thedegradation of certain compounds.

Anhydrous pharmaceutical compositions and dosage forms of the inventioncan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. An anhydrous pharmaceuticalcomposition may be prepared and stored such that its anhydrous nature ismaintained. Accordingly, anhydrous compositions are packaged usingmaterials known to prevent exposure to water such that they can beincluded in suitable formulary kits. Examples of suitable packaginginclude, but are not limited to, hermetically sealed foils, plastics,unit dose containers (e.g., vials), blister packs, and strip packs.

The invention further provides pharmaceutical compositions and dosageforms that comprise one or more agents that reduce the rate by which thecompound of the present invention as an active ingredient willdecompose. Such agents, which are referred to herein as “stabilizers,”include, but are not limited to, antioxidants such as ascorbic acid, pHbuffers, or salt buffers, etc.

The compounds of formula I in free form or in salt form, exhibitvaluable pharmacological properties, e.g. PI3K modulating properties,e.g. as indicated in in vitro and in vivo tests as provided in the nextsections, and are therefore indicated for therapy or for use as researchchemicals, e.g. as tool compounds.

Compounds of the invention may be useful in the treatment of conditions,diseases or disorders including disease or infection associatedimmunopathology in which one or more of the functions of B cells such asantibody production, antigen presentation, cytokine production orlymphoid organogenesis are abnormal or are undesirable includingrheumatoid arthritis, pemphigus vulgaris and related diseases,idiopathic thrombocytopenia purpura, systemic lupus erythematosus,multiple sclerosis, myasthenia gravis, Sjögren's syndrome, autoimmunehemolytic anemia, ANCA-associated vasculitides, cryoglobulinemia,thrombotic thrombocytopenic purpura, chronic autoimmune urticaria,allergy (atopic dermatitis, contact dermatitis, allergic rhinitis),goodpasture's syndrome, AMR (antibody-mediated transplant rejection), Bcell-mediated hyperacute, acute and chronic transplant rejection andcancers of haematopoietic origin including but not limited to multiplemyeloma; acute myelogenous leukemia; chronic myelogenous leukemia;lymphocytic leukemia; myeloid leukemia; non-Hodgkin lymphoma; lymphomas;polycythemia vera; essential thrombocythemia; myelofibrosis with myeloidmetaplasia; and Walden stroem disease.

The invention includes methods of treating conditions, diseases ordisorders in which one or more of the functions of neutrophils, such assuperoxide release, stimulated exocytosis, or chemoatractic migrationare abnormal or are undesirable including rheumatoid arthritis, sepsis,pulmonary or respiratory disorders such as asthma, inflammatorydermatoses such as psoriasis, as well as in disease or infectionassociated immunopathology and others.

The invention includes methods of treating conditions, diseases ordisorders in which one or more of the functions of basophil and mastcells such as chemoatractic migration or allergen-IgE-mediateddegranulation are abnormal or are undesirable including allergicdiseases (atopic dermatitis, contact dermatitis, allergic rhinitis) aswell as other disorders such as COPD, asthma or emphysema.

The invention includes methods of treating conditions, diseases ordisorders in which one or more of the functions of T cells such ascytokine production or cell-mediated cytotoxicity abnormal or areundesirable including rheumatoid arthritis, multiple sclerosis, acute orchronic rejection of cell tissue or organ grafts or cancers ofhaematopoietic origin as well as in disease or infection associatedimmunopathology.

Further, the invention includes methods of treating neurodegenerativediseases, cardiovascular diseases and platelet aggregation.

Further, the invention includes methods of treating skin diseases suchas porphyria cutanea tarda, polymorphous light eruption,dermatomyositis, solar urticaria, oral lichen planus, panniculitis,scleroderma, urticarial vasculitis.

Further, the invention includes methods of treating chronic inflammatorydiseases such as sarcoidosis, granuloma annulare.

In other embodiments, the condition or disorder (e.g. PI3K-mediated) isselected from the group consisting of: polycythemia vera, essentialthrombocythemia, myelofibrosis with myeloid metaplasia, asthma, COPD,ARDS, Loffler's syndrome, eosinophilic pneumonia, parasitic (inparticular metazoan) infestation (including tropical eosinophilia),bronchopulmonary aspergillosis, polyarteritis nodosa (includingChurg-Strauss syndrome), eosinophilic granuloma, eosinophil-relateddisorders affecting the airways occasioned by drug-reaction, psoriasis,contact dermatitis, atopic dermatitis, alopecia areata, erythemamultiforme, dermatitis herpetiformis, scleroderma, vitiligo,hypersensitivity angiitis, urticaria, bullous pemphigoid, lupuserythematosus, pemphigus, epidermolysis bullosa acquisita, autoimmunehaematogical disorders (e.g. haemolytic anaemia, aplastic anaemia, purered cell anaemia and idiopathic thrombocytopenia), systemic lupuserythematosus, polychondritis, scleroderma, Wegener granulomatosis,dermatomyositis, chronic active hepatitis, myasthenia gravis,Steven-Johnson syndrome, idiopathic sprue, autoimmune inflammatory boweldisease (e.g. ulcerative colitis and Crohn's disease), endocrineopthalmopathy, Grave's disease, sarcoidosis, alveolitis, chronichypersensitivity pneumonitis, multiple sclerosis, primary biliarycirrhosis, uveitis (anterior and posterior), interstitial lung fibrosis,psoriatic arthritis, glomerulonephritis, cardiovascular diseases,atherosclerosis, hypertension, deep venous thrombosis, stroke,myocardial infarction, unstable angina, thromboembolism, pulmonaryembolism, thrombolytic diseases, acute arterial ischemia, peripheralthrombotic occlusions, and coronary artery disease, reperfusioninjuries, retinopathy, such as diabetic retinopathy or hyperbaricoxygen-induced retinopathy, and conditions characterized by elevatedintraocular pressure or secretion of ocular aqueous humor, such asglaucoma.

In another embodiment, the compounds of the present invention are usefulin the treatment, prevention, or amelioration of autoimmune disease andof inflammatory conditions, in particular inflammatory conditions withan aetiology including an autoimmune component such as arthritis (forexample rheumatoid arthritis, arthritis chronica progrediente andarthritis deformans) and rheumatic diseases, including inflammatoryconditions and rheumatic diseases involving bone loss, inflammatorypain, spondyloarhropathies including ankolsing spondylitis, Reitersyndrome, reactive arthritis, psoriatic arthritis, and enterophathicsarthritis, hypersensitivity (including both airways hypersensitivity anddermal hypersensitivity) and allergies. Specific auto-immune diseasesfor which antibodies of the invention may be employed include autoimmunehaematological disorders (including e.g. hemolytic anaemia, aplasticanaemia, pure red cell anaemia and idiopa-thic thrombocytopenia),acquired hemophilia A, cold agglutinin disease, cryoglobulinemia,thrombotic thrombocytopenic purpura, Sjögren's syndrome, systemic lupuserythematosus, inflammatory muscle disorders, polychondritis,sclerodoma, anti-neutrophil cytoplasmic antibody-associated vasculitis,IgM mediated neuropathy, opsoclonus myoclonus syndrome, Wegenergranulomatosis, dermatomyositis, chronic active hepatitis, myastheniagravis, psoriasis, Steven-Johnson syndrome, pemphigus vulgaris,pemphigus foliacius, idio-pathic sprue, autoimmune inflammatory boweldisease (including e.g. ulcerative colitis, Crohn's disease andIrritable Bowel Syndrome), endocrine ophthalmopathy, Graves' disease,sarcoidosis, multiple sclerosis, neuromyelitis optica, primary biliarycirrhosis, juvenile diabetes (diabetes mellitus type I), uveitis(anterior, intermediate and posterior as well as panuveitis),keratoconjunctivitis sicca and vernal keratoconjunctivitis, interstitiallung fibrosis, psoriatic arthritis and glomerulonephritis (with andwithout nephrotic syndrome, e.g. including idiopathic nephro-ticsyndrome or minimal change nephropathy), tumors, inflammatory disease ofskin and cornea, myositis, loosening of bone implants, metabolicdisorders, such as atherosclerosis, diabetes, and dislipidemia.

In another embodiment, the compounds of the present invention are usefulin the treatment of conditions or disorders selected from the groupconsisting of, primary cutaneous B-cell lymphoma, immunobullous disease,pemphigus vulgaris, pemphigus foliaceus, endemic form of Brazilianpemphigus (Fogo selvagem), paraneoplastic pemphigus, bullous pemphigoid,mucous membrane pemphigoid, epidermolysis bullosa acquisita, chronicgraft versus host disease, dermatomyositis, systemic lupuserythematosus, vasculitis, small vessel vasculitis, hypocomplementemicurticarial vasculitis, antineutrophil cytoplasmic antibody-vasculitis,cryoglobulinemia, Schnitzler syndrome, Waldenstrom's macroglobulinemia,angioedema, vitiligo, systemic lupus erythematosus, idiopathicthrombocytopenic purpura, multiple sclerosis, cold agglutinin disease,autoimmune hemolytic anemia, antineutrophil cytoplasmicantibody-associated vasculitis, graft versus host disease,cryoglobulinemia and thrombotic thrombocytopenic.

Thus, as a further embodiment, the present invention provides the use ofa compound of formulae (I), (I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′),(Id), (Id′), (Ie), (Ie′), (If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii),(Ii′), (Ij) or (Ij′) in therapy. In a further embodiment, the therapy isselected from a disease which may be treated by inhibition of PI3K. Inanother embodiment, the disease is selected from the afore-mentionedlist, suitably from autoimmune disorders, inflammatory diseases,allergic diseases, airway diseases, such as asthma and COPD, transplantrejection; antibody production, antigen presentation, cytokineproduction or lymphoid organogenesis are abnormal or are undesirableincluding rheumatoid arthritis, pemphigus vulgaris, idiopathicthrombocytopenia purpura, systemic lupus erythematosus, multiplesclerosis, myasthenia gravis, Sjögren's syndrome, autoimmune hemolyticanemia, ANCA-associated vasculitides, cryoglobulinemia, thromboticthrombocytopenic purpura, chronic autoimmune urticaria, allergy (atopicdermatitis, contact dermatitis, allergic rhinitis), goodpasture'ssyndrome, AMR (antibody-mediated transplant rejection), B cell-mediatedhyperacute, acute and chronic transplant rejection and cancers ofhaematopoietic origin including but not limited to multiple myeloma; aleukaemia; acute myelogenous leukemia; chronic myelogenous leukemia;lymphocytic leukemia; myeloid leukemia; non-Hodgkin lymphoma; lymphomas;polycythemia vera; essential thrombocythemia; myelofibrosis with myeloidmetaplasia; and Walden stroem disease; more suitably from rheumatoidarthritis (RA), pemphigus vulgaris (PV), idiopathic thrombocytopeniapurpura (ITP), thrombotic thrombocytopenic purpura (TTP), autoimmunehemolytic anemia (AIHA), acquired hemophilia type A (AHA), systemiclupus erythematosus (SLE), multiple sclerosis (MS), myasthenia gravis(MG), Sjögren's syndrome (SS), ANCA-associated vasculitides,cryoglobulinemia, chronic autoimmune urticaria (CAU), allergy (atopicdermatitis, contact dermatitis, allergic rhinitis), goodpasture'ssyndrome, transplant rejection and cancers of haematopoietic origin aswell as in disease or infection associated immunopathology, for examplein severe and cerebral malaria, trypanosomiasis, leishmaniasis,toxoplasmosis and neurocysticercosis.

In another embodiment, the invention provides a method of treating adisease which is treated by inhibition of PI3K comprising administrationof a therapeutically acceptable amount of a compound of formulae (I),(I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie), (Ie′),(If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii), (Ii′), (Ij) or (Ij′). In afurther embodiment, the disease is selected from the afore-mentionedlist, suitably from autoimmune disorders, inflammatory diseases,allergic diseases, airway diseases, such as asthma and COPD, transplantrejection; antibody production, antigen presentation, cytokineproduction or lymphoid organogenesis are abnormal or are undesirableincluding rheumatoid arthritis, pemphigus vulgaris, idiopathicthrombocytopenia purpura, systemic lupus erythematosus, multiplesclerosis, myasthenia gravis, Sjögren's syndrome, autoimmune hemolyticanemia, ANCA-associated vasculitides, cryoglobulinemia, thromboticthrombocytopenic purpura, chronic autoimmune urticaria, allergy (atopicdermatitis, contact dermatitis, allergic rhinitis), goodpasture'ssyndrome, AMR (antibody-mediated transplant rejection), B cell-mediatedhyperacute, acute and chronic transplant rejection and cancers ofhaematopoietic origin including but not limited to multiple myeloma; aleukaemia; acute myelogenous leukemia; chronic myelogenous leukemia;lymphocytic leukemia; myeloid leukemia; non-Hodgkin lymphoma; lymphomas;polycythemia vera; essential thrombocythemia; myelofibrosis with myeloidmetaplasia; and Walden stroem disease; more suitably from rheumatoidarthritis (RA), pemphigus vulgaris (PV), idiopathic thrombocytopeniapurpura (ITP), thrombotic thrombocytopenic purpura (TTP), autoimmunehemolytic anemia (AIHA), acquired hemophilia type A (AHA), systemiclupus erythematosus (SLE), multiple sclerosis (MS), myasthenia gravis(MG), Sjögren's syndrome (SS), ANCA-associated vasculitides,cryoglobulinemia, chronic autoimmune urticaria (CAU), allergy (atopicdermatitis, contact dermatitis, allergic rhinitis), goodpasture'ssyndrome, transplant rejection and cancers of haematopoietic origin aswell as in disease or infection associated immunopathology, for examplein severe and cerebral malaria, trypanosomiasis, leishmaniasis,toxoplasmosis and neurocysticercosis.

Thus, as a further embodiment, the present invention provides the use ofa compound of formulae (I), (I′), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′),(Id), (Id′), (Ie), (Ie′), (If), (If′), (Ig), (Ig′), (Ih), (Ih′), (Ii),(Ii′), (Ij) or (Ij′) for the manufacture of a medicament. In a furtherembodiment, the medicament is for treatment of a disease which may betreated inhibition of PI3K. In another embodiment, the disease isselected from the afore-mentioned list, suitably from autoimmunedisorders, inflammatory diseases, allergic diseases, airway diseases,such as asthma and COPD, transplant rejection; antibody production,antigen presentation, cytokine production or lymphoid organogenesis areabnormal or are undesirable including rheumatoid arthritis, pemphigusvulgaris, idiopathic thrombocytopenia purpura, systemic lupuserythematosus, multiple sclerosis, myasthenia gravis, Sjögren'ssyndrome, autoimmune hemolytic anemia, ANCA-associated vasculitides,cryoglobulinemia, thrombotic thrombocytopenic purpura, chronicautoimmune urticaria, allergy (atopic dermatitis, contact dermatitis,allergic rhinitis), goodpasture's syndrome, AMR (antibody-mediatedtransplant rejection), B cell-mediated hyperacute, acute and chronictransplant rejection and cancers of haematopoietic origin including butnot limited to multiple myeloma; a leukaemia; acute myelogenousleukemia; chronic myelogenous leukemia; lymphocytic leukemia; myeloidleukemia; non-Hodgkin lymphoma; lymphomas; polycythemia vera; essentialthrombocythemia; myelofibrosis with myeloid metaplasia; and Waldenstroem disease; more suitably from rheumatoid arthritis (RA), pemphigusvulgaris (PV), idiopathic thrombocytopenia purpura (ITP), thromboticthrombocytopenic purpura (TTP), autoimmune hemolytic anemia (AIHA),acquired hemophilia type A (AHA), systemic lupus erythematosus (SLE),multiple sclerosis (MS), myasthenia gravis (MG), Sjögren's syndrome(SS), ANCA-associated vasculitides, cryoglobulinemia, chronic autoimmuneurticaria (CAU), allergy (atopic dermatitis, contact dermatitis,allergic rhinitis), goodpasture's syndrome, transplant rejection andcancers of haematopoietic origin as well as in disease or infectionassociated immunopathology, for example in severe and cerebral malaria,trypanosomiasis, leishmaniasis, toxoplasmosis and neurocysticercosis.

The pharmaceutical composition or combination of the present inventioncan be in unit dosage of about 1-1000 mg of active ingredient(s) for asubject of about 50-70 kg, or about 1-500 mg or about 1-250 mg or about1-150 mg or about 0.5-100 mg, or about 1-50 mg of active ingredients.The therapeutically effective dosage of a compound, the pharmaceuticalcomposition, or the combinations thereof, is dependent on the species ofthe subject, the body weight, age and individual condition, the disorderor disease or the severity thereof being treated. A physician, clinicianor veterinarian of ordinary skill can readily determine the effectiveamount of each of the active ingredients necessary to prevent, treat orinhibit the progress of the disorder or disease.

The above-cited dosage properties are demonstrable in vitro and in vivotests using advantageously mammals, e.g., mice, rats, dogs, monkeys orisolated organs, tissues and preparations thereof. The compounds of thepresent invention can be applied in vitro in the form of solutions,e.g., aqueous solutions, and in vivo either enterally, parenterally,advantageously intravenously, e.g., as a suspension or in aqueoussolution. The dosage in vitro may range between about 10⁻³ molar and10⁻⁹ molar concentrations. A therapeutically effective amount in vivomay range depending on the route of administration, between about0.1-500 mg/kg, or between about 1-100 mg/kg.

The compound of the present invention may be administered eithersimultaneously with, or before or after, one or more other therapeuticagent. The compound of the present invention may be administeredseparately, by the same or different route of administration, ortogether in the same pharmaceutical composition as the other agents.

In one embodiment, the invention provides a product comprising acompound of formula (I) and at least one other therapeutic agent as acombined preparation for simultaneous, separate or sequential use intherapy. In one embodiment, the therapy is the treatment of a disease orcondition mediated by the activity of the PI3K enzymes. Productsprovided as a combined preparation include a composition comprising thecompound of formula (I) and the other therapeutic agent(s) together inthe same pharmaceutical composition, or the compound of formula (I) andthe other therapeutic agent(s) in separate form, e.g. in the form of akit.

In one embodiment, the invention provides a pharmaceutical compositioncomprising a compound of formula (I) and another therapeutic agent(s).Optionally, the pharmaceutical composition may comprise apharmaceutically acceptable carrier, as described above.

In one embodiment, the invention provides a kit comprising two or moreseparate pharmaceutical compositions, at least one of which contains acompound of formula (I). In one embodiment, the kit comprises means forseparately retaining said compositions, such as a container, dividedbottle, or divided foil packet. An example of such a kit is a blisterpack, as typically used for the packaging of tablets, capsules and thelike.

The kit of the invention may be used for administering different dosageforms, for example, oral and parenteral, for administering the separatecompositions at different dosage intervals, or for titrating theseparate compositions against one another. To assist compliance, the kitof the invention typically comprises directions for administration.

In the combination therapies of the invention, the compound of theinvention and the other therapeutic agent may be manufactured and/orformulated by the same or different manufacturers. Moreover, thecompound of the invention and the other therapeutic may be broughttogether into a combination therapy: (i) prior to release of thecombination product to physicians (e.g. in the case of a kit comprisingthe compound of the invention and the other therapeutic agent); (ii) bythe physician themselves (or under the guidance of the physician)shortly before administration; (iii) in the patient themselves, e.g.during sequential administration of the compound of the invention andthe other therapeutic agent.

Accordingly, the invention provides the use of a compound of formula (I)for treating a disease or condition mediated by the activity of the PI3Kenzymes, wherein the medicament is prepared for administration withanother therapeutic agent. The invention also provides the use ofanother therapeutic agent for treating a disease or condition mediatedby the activity of the PI3K enzymes, wherein the medicament isadministered with a compound of formula (I).

The invention also provides a compound of formula (I) for use in amethod of treating a disease or condition mediated by the activity ofthe PI3K enzymes, wherein the compound of formula (I) is prepared foradministration with another therapeutic agent. The invention alsoprovides another therapeutic agent for use in a method of treating adisease or condition mediated by the activity of the PI3K enzymes,wherein the other therapeutic agent is prepared for administration witha compound of formula (I). The invention also provides a compound offormula (I) for use in a method of treating a disease or conditionmediated by the activity of the PI3K enzymes wherein the compound offormula (I) is administered with another therapeutic agent. Theinvention also provides another therapeutic agent for use in a method oftreating a disease or condition mediated by the activity of the PI3Kenzymes wherein the other therapeutic agent is administered with acompound of formula (I).

The invention also provides the use of a compound of formula (I) fortreating a disease or condition mediated by the activity of the PI3Kenzymes, wherein the patient has previously (e.g. within 24 hours) beentreated with another therapeutic agent. The invention also provides theuse of another therapeutic agent for treating a disease or conditionmediated by the activity of the PI3K enzymes, wherein the patient haspreviously (e.g. within 24 hours) been treated with a compound offormula (I).

The compounds of formula I may be administered as the sole activeingredient or in conjunction with, e.g. as an adjuvant to, other drugse.g. immunosuppressive or immunomodulating agents or otheranti-inflammatory agents, e.g. for the treatment or prevention of allo-or xenograft acute or chronic rejection or inflammatory or autoimmunedisorders, or a chemotherapeutic agent, e.g. a malignant cellanti-proliferative agent. For example, the compounds of formula I may beused in combination with a calcineurin inhibitor, e.g. cyclosporin A orFK 506; a mTOR inhibitor, e.g. rapamycin,40-O-(2-hydroxyethyl)-rapamycin, CCI1779, ABT578, AP23573, TAFA-93,biolimus-7 or biolimus-9; an ascomycin having immuno-suppressiveproperties, e.g. ABT-281, ASM981, etc.; corticosteroids;cyclophosphamide; azathioprene; methotrexate; leflunomide; mizoribine;mycophenolic acid or salt; mycophenolate mofetil; 15-deoxyspergualine oran immunosuppressive homologue, analogue or derivative thereof; a PKCinhibitor, e.g. as disclosed in WO 02/38561 or WO 03/82859, e.g. thecompound of Example 56 or 70; a JAK3 kinase inhibitor, e.g.N-benzyl-3,4-dihydroxy-benzylidene-cyanoacetamideα-cyano-(3,4-dihydroxy)-]N-benzylcinnamamide (Tyrphostin AG 490),prodigiosin 25-C (PNU156804),[4-(4′-hydroxyphenyl)-amino-6,7-dimethoxyquinazoline] (WHI-P131),[4-(3′-bromo-4′-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline](WHI-P154),[4-(3′,5′-dibromo-4′-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline]WHI-P97, KRX-211,3-{(3R,4R)-4-methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile,in free form or in a pharmaceutically acceptable salt form, e.g.mono-citrate (also called CP-690,550), or a compound as disclosed in WO04/052359 or WO 05/066156; immunosuppressive monoclonal antibodies,e.g., monoclonal antibodies to leukocyte receptors, e.g., MHC, CD2, CD3,CD4, CD7, CD8, CD25, CD28, CD40, CD45, CD52, CD58, CD80, CD86 or theirligands; other immunomodulatory compounds, e.g. a recombinant bindingmolecule having at least a portion of the extracellular domain of CTLA4or a mutant thereof, e.g. an at least extracellular portion of CTLA4 ora mutant thereof joined to a non-CTLA4 protein sequence, e.g. CTLA4Ig(for ex. designated ATCC 68629) or a mutant thereof, e.g. LEA29Y;adhesion molecule inhibitors, e.g. LFA-1 antagonists, ICAM-1 or -3antagonists, VCAM-4 antagonists or VLA-4 antagonists; or antihistamines;or antitussives, or a bronchodilatory agent; or an angiotensin receptorblockers; or an anti-infectious agent.

Where the compounds of formula I are administered in conjunction withother immunosuppressive/immunomodulatory, anti-inflammatory,chemotherapeutic or anti-infectious therapy, dosages of theco-administered immunosuppressant, immunomodulatory, anti-inflammatory,chemotherapeutic or anti-infectious compound will of course varydepending on the type of co-drug employed, e.g. whether it is a steroidor a calcineurin inhibitor, on the specific drug employed, on thecondition being treated and so forth.

A compound of the formula (I) may also be used to advantage incombination with each other or in combination with other therapeuticagents, especially other antiproliferative agents. Suchantiproliferative agents include, but are not limited to, aromataseinhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase IIinhibitors; microtubule active agents; alkylating agents; histonedeacetylase inhibitors; compounds, which induce cell differentiationprocesses; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors;antineoplastic antimetabolites; platin compounds; compoundstargeting/decreasing a protein or lipid kinase activity and furtheranti-angiogenic compounds; compounds which target, decrease or inhibitthe activity of a protein or lipid phosphatase; gonadorelin agonists;anti-androgens; methionine aminopeptidase inhibitors; bisphosphonates;biological response modifiers; antiproliferative antibodies; heparanaseinhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors;proteasome inhibitors; agents used in the treatment of hematologicmalignancies; compounds which target, decrease or inhibit the activityof Flt-3; Hsp90 inhibitors; temozolomide (TEMODAL®); and leucovorin.

The term “aromatase inhibitor”, as used herein, relates to a compoundwhich inhibits the estrogen production, i.e., the conversion of thesubstrates androstenedione and testosterone to estrone and estradiol,respectively. The term includes, but is not limited to, steroids,especially atamestane, exemestane and formestane; and, in particular,non-steroids, especially aminoglutethimide, roglethimide,pyridoglutethimide, trilostane, testolactone, ketoconazole, vorozole,fadrozole, anastrozole and letrozole. Exemestane can be administered,e.g., in the form as it is marketed, e.g., under the trademark AROMASIN.Formestane can be administered, e.g., in the form as it is marketed,e.g., under the trademark LENTARON. Fadrozole can be administered, e.g.,in the form as it is marketed, e.g., under the trademark AFEMA.Anastrozole can be administered, e.g., in the form as it is marketed,e.g., under the trademark ARIMIDEX. Letrozole can be administered, e.g.,in the form as it is marketed, e.g., under the trademark FEMARA orFEMAR. Aminoglutethimide can be administered, e.g., in the form as it ismarketed, e.g., under the trademark ORIMETEN. A combination of theinvention comprising a chemotherapeutic agent which is an aromataseinhibitor is particularly useful for the treatment of hormone receptorpositive tumors, e.g., breast tumors.

The term “anti-estrogen”, as used herein, relates to a compound whichantagonizes the effect of estrogens at the estrogen receptor level. Theterm includes, but is not limited to, tamoxifen, fulvestrant, raloxifeneand raloxifene hydrochloride. Tamoxifen can be administered, e.g., inthe form as it is marketed, e.g., under the trademark NOLVADEX.Raloxifene hydrochloride can be administered, e.g., in the form as it ismarketed, e.g., under the trademark EVISTA. Fulvestrant can beformulated as disclosed in U.S. Pat. No. 4,659,516 or it can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark FASLODEX. A combination of the invention comprising achemotherapeutic agent which is an antiestrogen is particularly usefulfor the treatment of estrogen receptor positive tumors, e.g., breasttumors.

The term “anti-androgen”, as used herein, relates to any substance whichis capable of inhibiting the biological effects of androgenic hormonesand includes, but is not limited to, bicalutamide (CASODEX), which canbe formulated, e.g., as disclosed in U.S. Pat. No. 4,636,505.

The term “gonadorelin agonist”, as used herein, includes, but is notlimited to, abarelix, goserelin and goserelin acetate. Goserelin isdisclosed in U.S. Pat. No. 4,100,274 and can be administered, e.g., inthe form as it is marketed, e.g., under the trademark ZOLADEX. Abarelixcan be formulated, e.g., as disclosed in U.S. Pat. No. 5,843,901.

The term “topoisomerase I inhibitor”, as used herein, includes, but isnot limited to, topotecan, gimatecan, irinotecan, camptothecian and itsanalogues, 9-nitrocamptothecin and the macromolecular camptothecinconjugate PNU-166148 (compound A1 in WO 99/17804). Irinotecan can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark CAMPTOSAR. Topotecan can be administered, e.g., in the form asit is marketed, e.g., under the trademark HYCAMTIN.

The term “topoisomerase II inhibitor”, as used herein, includes, but isnot limited to, the anthracyclines, such as doxorubicin, includingliposomal formulation, e.g., CAELYX; daunorubicin; epirubicin;idarubicin; nemorubicin; the anthraquinones mitoxantrone andlosoxantrone; and the podophillotoxines etoposide and teniposide.Etoposide can be administered, e.g., in the form as it is marketed,e.g., under the trademark ETOPOPHOS. Teniposide can be administered,e.g., in the form as it is marketed, e.g., under the trademark VM26-BRISTOL. Doxorubicin can be administered, e.g., in the form as it ismarketed, e.g., under the trademark ADRIBLASTIN or ADRIAMYCIN.Epirubicin can be administered, e.g., in the form as it is marketed,e.g., under the trademark FARMORUBICIN. Idarubicin can be administered,e.g., in the form as it is marketed, e.g., under the trademark ZAVEDOS.Mitoxantrone can be administered, e.g., in the form as it is marketed,e.g., under the trademark NOVANTRON.

The term “microtubule active agent” relates to microtubule stabilizing,microtubule destabilizing agents and microtublin polymerizationinhibitors including, but not limited to, taxanes, e.g., paclitaxel anddocetaxel; vinca alkaloids, e.g., vinblastine, especially vinblastinesulfate; vincristine, especially vincristine sulfate and vinorelbine;discodermolides; cochicine; and epothilones and derivatives thereof,e.g., epothilone B or D or derivatives thereof. Paclitaxel may beadministered, e.g., in the form as it is marketed, e.g., TAXOL.Docetaxel can be administered, e.g., in the form as it is marketed,e.g., under the trademark TAXOTERE. Vinblastine sulfate can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark VINBLASTIN R.P. Vincristine sulfate can be administered, e.g.,in the form as it is marketed, e.g., under the trademark FARMISTIN.Discodermolide can be obtained, e.g., as disclosed in U.S. Pat. No.5,010,099. Also included are epothilone derivatives which are disclosedin WO 98/10121, U.S. Pat. No. 6,194,181, WO 98/25929, WO 98/08849, WO99/43653, WO 98/22461 and WO 00/31247. Especially preferred areepothilone A and/or B.

The term “alkylating agent”, as used herein, includes, but is notlimited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNUor Gliadel). Cyclophosphamide can be administered, e.g., in the form asit is marketed, e.g., under the trademark CYCLOSTIN. Ifosfamide can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark HOLOXAN.

The term “histone deacetylase inhibitors” or “HDAC inhibitors” relatesto compounds which inhibit the histone deacetylase and which possessantiproliferative activity. This includes compounds disclosed in WO02/22577, especiallyN-hydroxy-3-[4-[[(2-hydroxyethyl)[2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide,N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamideand pharmaceutically acceptable salts thereof. It further especiallyincludes suberoylanilide hydroxamic acid (SAHA).

The term “antineoplastic antimetabolite” includes, but is not limitedto, 5-fluorouracil or 5-FU; capecitabine; gemcitabine; DNA demethylatingagents, such as 5-azacytidine and decitabine; methotrexate andedatrexate; and folic acid antagonists, such as pemetrexed. Capecitabinecan be administered, e.g., in the form as it is marketed, e.g., underthe trademark XELODA. Gemcitabine can be administered, e.g., in the formas it is marketed, e.g., under the trademark GEMZAR. Also included isthe monoclonal antibody trastuzumab which can be administered, e.g., inthe form as it is marketed, e.g., under the trademark HERCEPTIN.

The term “platin compound”, as used herein, includes, but is not limitedto, carboplatin, cis-platin, cisplatinum and oxaliplatin. Carboplatincan be administered, e.g., in the form as it is marketed, e.g., underthe trademark CARBOPLAT. Oxaliplatin can be administered, e.g., in theform as it is marketed, e.g., under the trademark ELOXATIN.

The term “compounds targeting/decreasing a protein or lipid kinaseactivity; or a protein or lipid phosphatase activity; or furtheranti-angiogenic compounds”, as used herein, includes, but is not limitedto, protein tyrosine kinase and/or serine and/or threonine kinaseinhibitors or lipid kinase inhibitors, e.g.,

-   -   a) compounds targeting, decreasing or inhibiting the activity of        the platelet-derived growth factor-receptors (PDGFR), such as        compounds which target, decrease or inhibit the activity of        PDGFR, especially compounds which inhibit the PDGF receptor,        e.g., a N-phenyl-2-pyrimidine-amine derivative, e.g., imatinib,        SU101, SU6668 and GFB-111;    -   b) compounds targeting, decreasing or inhibiting the activity of        the fibroblast growth factor-receptors (FGFR);    -   c) compounds targeting, decreasing or inhibiting the activity of        the insulin-like growth factor receptor I (IGF-IR), such as        compounds which target, decrease or inhibit the activity of        IGF-IR, especially compounds which inhibit the IGF-IR receptor,        such as those compounds disclosed in WO 02/092599;    -   d) compounds targeting, decreasing or inhibiting the activity of        the Trk receptor tyrosine kinase family;    -   e) compounds targeting, decreasing or inhibiting the activity of        the Axl receptor tyrosine kinase family;    -   f) compounds targeting, decreasing or inhibiting the activity of        the c-Met receptor;    -   g) compounds targeting, decreasing or inhibiting the activity of        the Kit/SCFR receptor tyrosine kinase;    -   h) compounds targeting, decreasing or inhibiting the activity of        the C-kit receptor tyrosine kinases—(part of the PDGFR family),        such as compounds which target, decrease or inhibit the activity        of the c-Kit receptor tyrosine kinase family, especially        compounds which inhibit the c-Kit receptor, e.g., imatinib;    -   i) compounds targeting, decreasing or inhibiting the activity of        members of the c-Abl family and their gene-fusion products,        e.g., BCR-Abl kinase, such as compounds which target decrease or        inhibit the activity of c-Abl family members and their gene        fusion products, e.g., a N-phenyl-2-pyrimidine-amine derivative,        e.g., imatinib, PD180970, AG957, NSC 680410 or PD173955 from        ParkeDavis;    -   j) compounds targeting, decreasing or inhibiting the activity of        members of the protein kinase C (PKC) and Raf family of        serine/threonine kinases, members of the MEK, SRC, JAK, FAK, PDK        and Ras/MAPK family members, or PI(3) kinase family, or of the        PI(3)-kinase-related kinase family, and/or members of the        cyclin-dependent kinase family (CDK) and are especially those        staurosporine derivatives disclosed in U.S. Pat. No. 5,093,330,        e.g., midostaurin; examples of further compounds include, e.g.,        UCN-01; safingol; BAY 43-9006; Bryostatin 1; Perifosine;        Ilmofosine; RO 318220 and RO 320432; GO 6976; Isis 3521;        LY333531/LY379196; isochinoline compounds, such as those        disclosed in WO 00/09495; FTIs; PD184352; or QAN697 (a PI3K        inhibitor);    -   k) compounds targeting, decreasing or inhibiting the activity of        protein-tyrosine kinase inhibitors, such as compounds which        target, decrease or inhibit the activity of protein-tyrosine        kinase inhibitors include imatinib mesylate (GLEEVEC) or        tyrphostin. A tyrphostin is preferably a low molecular weight        (Mr<1500) compound, or a pharmaceutically acceptable salt        thereof, especially a compound selected from the        benzylidenemalonitrile class or the S-arylbenzenemalonirile or        bisubstrate quinoline class of compounds, more especially any        compound selected from the group consisting of Tyrphostin        A23/RG-50810, AG 99, Tyrphostin AG 213, Tyrphostin AG 1748,        Tyrphostin AG 490, Tyrphostin B44, Tyrphostin B44 (+)        enantiomer, Tyrphostin AG 555, AG 494, Tyrphostin AG 556, AG957        and adaphostin (4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic        acid adamantyl ester, NSC 680410, adaphostin; and    -   l) compounds targeting, decreasing or inhibiting the activity of        the epidermal growth factor family of receptor tyrosine kinases        (EGFR, ErbB2, ErbB3, ErbB4 as homo- or hetero-dimers), such as        compounds which target, decrease or inhibit the activity of the        epidermal growth factor receptor family are especially        compounds, proteins or antibodies which inhibit members of the        EGF receptor tyrosine kinase family, e.g., EGF receptor, ErbB2,        ErbB3 and ErbB4 or bind to EGF or EGF related ligands, and are        in particular those compounds, proteins or monoclonal antibodies        generically and specifically disclosed in WO 97/02266, e.g., the        compound of Example 39, or in EP 0 564 409; WO 99/03854; EP        0520722; EP 0 566 226; EP 0 787 722; EP 0 837 063; U.S. Pat. No.        5,747,498; WO 98/10767; WO 97/30034; WO 97/49688; WO 97/38983        and, especially, WO 96/30347, e.g., compound known as CP 358774;        WO 96/33980, e.g., compound ZD 1839; and WO 95/03283, e.g.,        compound ZM105180, e.g., trastuzumab (HERCEPTIN), cetuximab,        Iressa, Tarceva, OSI-774, CI-1033, EKB-569, GW-2016, E1.1, E2.4,        E2.5, E6.2, E6.4, E2.11, E6.3 or E7.6.3; and        7H-pyrrolo-[2,3-d]pyrimidine derivatives which are disclosed in        WO 03/013541.

Further anti-angiogenic compounds include compounds having anothermechanism for their activity, e.g., unrelated to protein or lipid kinaseinhibition, e.g., thalidomide (THALOMID) and TNP-470.

Compounds which target, decrease or inhibit the activity of a protein orlipid phosphatase are, e.g., inhibitors of phosphatase 1, phosphatase2A, PTEN or CDC25, e.g., okadaic acid or a derivative thereof.

Compounds which induce cell differentiation processes are e.g. retinoicacid, α- γ- or δ-tocopherol or α- γ- or δ-tocotrienol.

The term cyclooxygenase inhibitor, as used herein, includes, but is notlimited to, e.g., Cox-2 inhibitors, 5-alkyl substituted2-arylaminophenylacetic acid and derivatives, such as celecoxib(CELEBREX), rofecoxib (VIOXX), etoricoxib, valdecoxib or a5-alkyl-2-arylaminophenylacetic acid, e.g.,5-methyl-2-(2′-chloro-6′-fluoroanilino)phenyl acetic acid orlumiracoxib.

The term “bisphosphonates”, as used herein, includes, but is not limitedto, etridonic, clodronic, tiludronic, pamidronic, alendronic,ibandronic, risedronic and zoledronic acid. “Etridonic acid” can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark DIDRONEL. “Clodronic acid” can be administered, e.g., in theform as it is marketed, e.g., under the trademark BONEFOS. “Tiludronicacid” can be administered, e.g., in the form as it is marketed, e.g.,under the trademark SKELID. “Pamidronic acid” can be administered, e.g.,in the form as it is marketed, e.g., under the trademark AREDIA™.“Alendronic acid” can be administered, e.g., in the form as it ismarketed, e.g., under the trademark FOSAMAX. “lIbandronic acid” can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark BONDRANAT. “Risedronic acid” can be administered, e.g., in theform as it is marketed, e.g., under the trademark ACTONEL. “Zoledronicacid” can be administered, e.g., in the form as it is marketed, e.g.,under the trademark ZOMETA.

The term “mTOR inhibitors” relates to compounds which inhibit themammalian target of rapamycin (mTOR) and which possess antiproliferativeactivity, such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779and ABT578.

The term “heparanase inhibitor”, as used herein, refers to compoundswhich target, decrease or inhibit heparin sulphate degradation. The termincludes, but is not limited to, PI-88.

The term “biological response modifier”, as used herein, refers to alymphokine or interferons, e.g., interferon γ.

The term “inhibitor of Ras oncogenic isoforms”, e.g., H-Ras, K-Ras orN-Ras, as used herein, refers to compounds which target, decrease orinhibit the oncogenic activity of Ras, e.g., a “farnesyl transferaseinhibitor”, e.g., L-744832, DK8G557 or R115777 (Zarnestra).

The term “telomerase inhibitor”, as used herein, refers to compoundswhich target, decrease or inhibit the activity of telomerase. Compoundswhich target, decrease or inhibit the activity of telomerase areespecially compounds which inhibit the telomerase receptor, e.g.,telomestatin.

The term “methionine aminopeptidase inhibitor”, as used herein, refersto compounds which target, decrease or inhibit the activity ofmethionine aminopeptidase. Compounds which target, decrease or inhibitthe activity of methionine aminopeptidase are, e.g., bengamide or aderivative thereof.

The term “proteasome inhibitor”, as used herein, refers to compoundswhich target, decrease or inhibit the activity of the proteasome.Compounds which target, decrease or inhibit the activity of theproteasome include, e.g., PS-341 and MLN 341.

The term “matrix metalloproteinase inhibitor” or “MMP inhibitor”, asused herein, includes, but is not limited to, collagen peptidomimeticand nonpeptidomimetic inhibitors, tetracycline derivatives, e.g.,hydroxamate peptidomimetic inhibitor batimastat and its orallybioavailable analogue marimastat (BB-2516), prinomastat (AG3340),metastat (NSC 683551) BMS-279251, BAY 12-9566, TAA211, MMI270B orAAJ996.

The term “agents used in the treatment of hematologic malignancies”, asused herein, includes, but is not limited to, FMS-like tyrosine kinaseinhibitors, e.g., compounds targeting, decreasing or inhibiting theactivity of FMS-like tyrosine kinase receptors (Flt-3R); interferon,1-b-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors,e.g., compounds which target, decrease or inhibit anaplastic lymphomakinase.

Compounds which target, decrease or inhibit the activity of FMS-liketyrosine kinase receptors (Flt-3R) are especially compounds, proteins orantibodies which inhibit members of the Flt-3R receptor kinase family,e.g., PKC412, midostaurin, a staurosporine derivative, SU11248 andMLN518.

The term “HSP90 inhibitors”, as used herein, includes, but is notlimited to, compounds targeting, decreasing or inhibiting the intrinsicATPase activity of HSP90; degrading, targeting, decreasing or inhibitingthe HSP90 client proteins via the ubiquitin proteasome pathway.Compounds targeting, decreasing or inhibiting the intrinsic ATPaseactivity of HSP90 are especially compounds, proteins or antibodies whichinhibit the ATPase activity of HSP90, e.g., 17-allylamino,17-demethoxygeldanamycin (17AAG), a geldanamycin derivative, othergeldanamycin related compounds, radicicol and HDAC inhibitors.

The term “antiproliferative antibodies”, as used herein, includes, butis not limited to, trastuzumab (Herceptin™), Trastuzumab-DM1, erlotinib(Tarceva™), bevacizumab (Avastin™), rituximab (Rituxan), PRO64553(anti-CD40) and 2C4 antibody. By antibodies is meant, e.g., intactmonoclonal antibodies, polyclonal antibodies, multispecific antibodiesformed from at least two intact antibodies, and antibodies fragments solong as they exhibit the desired biological activity.

For the treatment of acute myeloid leukemia (AML), compounds of formula(I) can be used in combination with standard leukemia therapies,especially in combination with therapies used for the treatment of AML.In particular, compounds of formula (I) can be administered incombination with, e.g., farnesyl transferase inhibitors and/or otherdrugs useful for the treatment of AML, such as Daunorubicin, Adriamycin,Ara-C, VP-16, Teniposide, Mitoxantrone, Idarubicin, Carboplatinum andPKC412.

A compound of the formula (I) may also be used to advantage incombination with each other or in combination with other therapeuticagents, especially other anti-malarial agents. Such anti-malarial agentsinclude, but are not limited to proguanil, chlorproguanil, trimethoprim,chloroquine, mefloquine, lumefantrine, atovaquone,pyrimethamine-sulfadoxine, pyrimethamine-dapsone, halofantrine, quinine,quinidine, amodiaquine, amopyroquine, sulphonamides, artemisinin,arteflene, artemether, artesunate, primaquine, inhaled NO, L-arginine,Dipropylenetri-amine NONOate (NO donor), Rosiglitzone (PPARy agonist),activated charcoal, Erythropoietin, Levamisole, and pyronaridine.

A compound of the formula (I) may also be used to advantage incombination with each other or in combination with other therapeuticagents, such as used for the treatment of Leishmaniosis,Trypanosomiasis, Toxoplasmosis and Neurocysticercosis. Such agentsinclude, but are not limited to chloroquine sulfate,atovaquone-proguanil, artemether-lumefantrine, quinine-sulfate,artesunate, quinine, doxycycline, clindamycin, meglumine antimoniate,sodium stibogluconate, miltefosine, ketoconazole, pentamidine,amphotericin B (AmB), liposomal-AmB, paromomycine, eflornithine,nifurtimox, suramin, melarsoprol, prednisolone, benznidazole,sulfadiazine, pyrimethamine, clindamycin, trimetropim, sulfamethoxazole,azitromycin, atovaquone, dexamethasone, praziquantel, albendazole,beta-lactams, fluoroquinolones, macrolides, aminoglycosides,sulfadiazine and pyrimethamine.

The structure of the active agents identified by code nos., generic ortrade names may be taken from the actual edition of the standardcompendium “The Merck Index” or from databases, e.g., PatentsInternational, e.g., IMS World Publications.

The above-mentioned compounds, which can be used in combination with acompound of the formula (I), can be prepared and administered asdescribed in the art, such as in the documents cited above.

A compound of the formula (I) may also be used to advantage incombination with known therapeutic processes, e.g., the administrationof hormones or especially radiation.

A compound of formula (I) may in particular be used as aradiosensitizer, especially for the treatment of tumors which exhibitpoor sensitivity to radiotherapy.

By “combination”, there is meant either a fixed combination in onedosage unit form, or a kit of parts for the combined administrationwhere a compound of the formula (I) and a combination partner may beadministered independently at the same time or separately within timeintervals that especially allow that the combination partners show acooperative, e.g., synergistic, effect or any combination thereof. Theterms “co-administration” or “combined administration” or the like asutilized herein are meant to encompass administration of the selectedcombination partner to a single subject in need thereof (e.g. apatient), and are intended to include treatment regimens in which theagents are not necessarily administered by the same route ofadministration or at the same time. The term “pharmaceuticalcombination” as used herein means a product that results from the mixingor combining of more than one active ingredient and includes both fixedand non-fixed combinations of the active ingredients. The term “fixedcombination” means that the active ingredients, e.g. a compound offormula I and a combination partner, are both administered to a patientsimultaneously in the form of a single entity or dosage. The term“non-fixed combination” means that the active ingredients, e.g. acompound of formula (I) and a combination partner, are both administeredto a patient as separate entities either simultaneously, concurrently orsequentially with no specific time limits, wherein such administrationprovides therapeutically effective levels of the two compounds in thebody of the patient. The latter also applies to cocktail therapy, e.g.the administration of three or more active ingredients.

EXAMPLES Experimental Details

The following examples are intended to illustrate the invention and arenot to be construed as being limitations thereon. Temperatures are givenin degrees Celsius. If not mentioned otherwise, all evaporations areperformed under reduced pressure, typically between about 15 mm Hg and100 mm Hg (=20-133 mbar). The structure of final products, intermediatesand starting materials is confirmed by standard analytical methods,e.g., microanalysis and spectroscopic characteristics, e.g., MS, IR,NMR. Abbreviations used are those conventional in the art.

All starting materials, building blocks, reagents, acids, bases,dehydrating agents, solvents, and catalysts utilized to synthesis thecompounds of the present invention are either commercially available orcan be produced by organic synthesis methods known to one of ordinaryskill in the art (Houben-Weyl 4th Ed. 1952, Methods of OrganicSynthesis, Thieme, Volume 21). Further, the compounds of the presentinvention can be produced by organic synthesis methods known to one ofordinary skill in the art as shown in the following examples.

ABBREVIATIONS

-   ACN acetonitrile-   AcOH acetic acid-   aq. aqueous-   Boc tert-butoxycarbonyl-   Boc₂O di-tert-butyl dicarbonate-   tBu tert-butyl-   tBuOH tert-butanol-   BrettPhos    2-(Dicyclohexylphosphino)-3,6-dimethoxy-2′-4′-6′-triisopropyl-1,1′-biphenyl-   br s broad singlet-   COMU    (1-cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbenium    hexafluorophosphate-   conc. concentrated-   d day(s)-   d doublet-   dd doublet of doublets-   dba dibenzylideneacetone-   DCM dichloromethane-   DEA diethylamine-   DEAD diethyl azodicarboxylate-   DEAP diethylaminopyridine-   DIPEA diisopropylethylamine-   DMF dimethylformamide-   DMME dimethoxymethane-   DMSO dimethylsulfoxide-   DPPA diphenylphosphoryl azide-   DPPF 1,1′-bis(diphenylphosphino)ferrocene-   EDC 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-   eq. equivalent(s)-   ESI electrospray ionisation-   Et₃N triethylamine-   Et₂O diethylether-   EtOAc ethyl acetate-   EtOH ethanol-   h hour(s)-   HATU O-(7-azabenzotriazol-1-yl)-N,N,NN′,N′-tetramethyluronium    hexafluorophosphate-   HBTU O-(1H-benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate-   HMDS hexamethyldisilazane-   HOBT 1-hydroxy-benztriazole-   HPLC high performance liquid chromatography-   IPA isopropanol-   LCMS liquid chromatography with mass spectrometry-   mCPBA meta-chloroperoxybenzoic acid-   MeOH methanol-   m multiplet-   min minute(s)-   MS mass spectrometry-   mw microwave-   NMR nuclear magnetic resonance spectrometry-   NaOtBu sodium tert-butoxide-   NP normal phase-   OBD optimum bed density-   Pd₂(dba)₃ tris(dibenzylideneacetone)dipalladium-   PL-HCO₃ MP SPE Polymer-supported bicarbonate cartridge for acid    removal-   prep. preparative-   PPh₃ triphenylphosphine-   q quartet-   Rac-BINAP racemic 2,2′-bis(di-p-tolylphosphino)-1,1′-binaphthyl-   RP reversed phase-   Rt retention time-   rt room temperature-   RuPhos 2-dicyclohexylphosphino-2′,6′-di-isopropoxy-1,1′-biphenyl-   sat. saturated-   SCX-2 polymer supported sulfonic acid macroporous polystyrene-   soln. solution-   t triplet-   TBME tert-butyl methyl ether-   TBAF tetrabutylammonium fluoride-   TBDMSCl tert-butyldimethylsilylchloride-   Tetramethyl-t-butyl-   XPhos    2-di-t-butylphosphino-3,4,5,6-tetramethyl-2′,4′,6′-triisopropylbiphenyl-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   TLC thin layer chromatography-   UPLC ultra performance liquid chromatography-   XPhos 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl-   Pd[RuPhos] (2-dicyclohexylphosphino-2′6′-diisopropyl-1    1′-biphenyl)(2-(2-aminoethyl)phenyl)palladium(II)

Microwave equipment used is a Biotage Initiator®

All compounds are named using AutoNom.

Preparation of Examples General Procedures

a) (R)-pyrrolidin-3-ol and an acid chloride of formula R⁴C(O)Cl orcarboxylic acid of formula R⁴C(O)OH were reacted to prepare an amide ofgeneral formula II. Those skilled in the art will appreciate that thereare many known ways of preparing amides. For example, see Mantalbetti,C. A. G. N and Falque, V., Amide bond formation and peptide coupling,Tetrahedron, 2005, 61(46), pp 10827-10852 and references cited therein.The following general methods i-ii have been used.

i. A soln. of the carboxylic acid and DMF (1 eq.) in DCM was treatedwith oxalyl chloride (1.5 eq.) for 1 h at 3° C. The reaction mixture wasconcentrated under reduced pressure, dissolved in DCM and added to asoln. of (R)-pyrrolidin-3-ol hydrochloride (1.0 eq.) and Et₃N (2.5 eq.)in DCM at 3° C. The resulting mixture was stirred vigorously at 3° C.for 1 h, then concentrated under reduced pressure. The residue wastreated with EtOAc and filtered. The residue was washed with EtOAc, andthe combined filtrates were concentrated under reduced pressure andpurified by flash chromatography.

ii. A soln. of a commercial acid chloride (1.0 eq.) in DCM was added toa soln. of (R)-pyrrolidin-3-ol hydrochloride (1.0 eq.) and Et₃N (2.5eq.) in DCM at 3° C. The resulting mixture was stirred vigorously at 3°C. for 1 h, then concentrated under reduced pressure. The residue wastreated with EtOAc and filtered. The residue was washed with EtOAc, andthe combined filtrates were concentrated under reduced pressure andpurified by flash chromatography. Typical conditions for amid bondformation reactions are exemplified in the section B) Amide bondformation conditions below.

b) The mesylates of compounds of general formula II were prepared bycustomary conditions, preferably by reaction of II with methane sulfonylchloride (2 eq.) and Et₃N (2 eq.) in DCM at 0° C.

c) Compounds of general formula V were prepared by reacting3,4-dihydro-2H-benzo[1,4]oxazin-6-ol IV with compounds of generalformula III in the presence of a suitable base such as sodium hydride(NaH) and polar organic solvent such as DMF under inert gas conditionsat 50° C. Typical conditions for such reactions are exemplified in thesection C) Side chain introduction conditions below.

d) Buchwald-Hartwig cross-coupling between V and an aryl halogenide ofthe general formula R²—X′ where X′=bromo or iodo was performed undercustomary Buchwald-Hartwig conditions using a Pd catalyst/ligandcombination such as preferablyPd₂(dba)₃/2-(dicyclohexylphosphino)biphenyl orPd₂(dba)₃/2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-biphenyl orbis(tri-t-butylphosphine)palladium and a base, such as preferablyNaOtBu, and organic solvent such as preferably toluene. The reaction waspreferably stirred at a temperature of approximately 80-120° C.,preferably 110° C. and was preferably performed in a microwave reactor.The reaction was preferably carried out under an inert gas such asnitrogen or argon. The final compounds were purified by normal orreversed phase chromatography. Typical conditions for Buchwald-Hartwigcross-coupling reactions are exemplified in the section A) Buchwaldaminations or hydroxylations below.

a) (S)-tert-butyl3-((3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)oxy)pyrrolidine-1-carboxylate(compound VIII) was prepared by reacting3,4-dihydro-2H-benzo[1,4]oxazin-6-ol IV with a compound of generalformula VII by one of the following methods 1) for X=mesylate, compoundsIV and VII were reacted in the presence of a suitable base such assodium hydride (NaH) and a polar organic solvent DMF under inert gasconditions at room temperature ii) for X=H, compounds of general formulaIV and VII were reacted using customary Mitsunobu conditions, preferablyusing Ph₃P (1.4 eq.) and DEAD (1.4 eq.) in organic solvent such as THFunder inert gas conditions at a temperature of preferably 70° C. Typicalconditions for such reactions are exemplified in the section C) Sidechain introduction conditions below.

b) Buchwald-Hartwig cross-coupling between VIII and an aryl halogenideof the general formula R²—X′ where X′=bromo or iodo was performed undercustomary Buchwald-Hartwig conditions using a Pd catalyst/ligandcombination such as preferably Pd₂(dba)₃/X-Phos, Pd₂(dba)₃/(rac)-BINAP,Pd(OAc)₂/(rac)-BINAP or bis(tri-t-butylphosphine)palladium and a base,such as preferably NaOtBu, Cs₂CO₃ or K₃PO₄ and an organic solvent suchas preferably toluene, dioxane or THF. The reaction was preferablystirred at a temperature of approximately 60-120° C. and was preferablybe performed in a microwave reactor. The reaction was preferably carriedout under an inert gas such as nitrogen or argon. Typical conditions forBuchwald-Hartwig cross-coupling reactions are exemplified in the sectionA) Buchwald aminations or hydroxylations below.

c) N—BOC deprotection of compounds of general formula IX was performedunder customary BOC deprotection conditions using among the possibleacids preferably trifluoro-acetic acid and organic solvent, preferablyDCM. The reaction was preferably performed at room temperature.

d) A compound of the general formula X and an acid chloride of formulaR⁴C(O)Cl or a carboxylic acid of formula R⁴C(O)OH were are reacted toprepare an amide of general formula VI using customary amide couplingconditions: in addition to the methods described in Scheme 1, step a)preferred coupling reagents were HBTU, HOBt/EDC, COMU/DIPEA. Thecouplings were performed in an organic solvent such as preferably DMF orDCM and the final compounds were purified by normal or reversed phasechromatography. Typical conditions for amid bond formation reactions areexemplified in the section B) Amide bond formation conditions below.

a) 3,4-Dihydro-2H-benzo[1,4]oxazin-6-ol IV was O-protected usingstandard silylation procedures, using a silylating reagent, preferablyTBDMSCl and a base, preferably NaH, in an organic solvent, preferablyTHF at room temperature.

b) Buchwald-Hartwig cross-coupling between XI and an aryl halogenide ofthe general formula R²—X′ where X′=bromo or iodo was performed undercustomary Buchwald-Hartwig conditions using a Pd catalyst/ligandcombination such as preferably Pd₂(dba)₃/X-Phos,Pd₂(dba)₃/dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl orbis(tri-t-butylphosphine)-palladium and a base, such as preferablyNaOtBu and an organic solvent such as preferably toluene. The reactionwas preferably stirred at a temperature of approximately 110-140° C. andwas preferably performed in a microwave reactor. The reaction waspreferably carried out under an inert gas such as nitrogen or argon.Typical conditions for Buchwald-Hartwig cross-coupling reactions areexemplified in the section A) Buchwald aminations or hydroxylationsbelow.

c) O-TBDMS deprotection of compounds of general formula XII wasperformed under customary deprotection conditions using preferably TBAFand an organic solvent, preferably THF. The reaction was preferablyperformed at room temperature

d) Compounds of general formula XIII were coupled with mesylates ofgeneral formula III using a suitable base such as preferably sodiumhydride (NaH) or K₂CO₃ and polar organic solvent such as DMF under inertgas conditions at room temperature or elevated temperatures up to 100°C. The final compounds were purified by normal or reversed phasechromatography. Typical conditions for such reactions are exemplified inthe section C) Side chain introduction conditions below.

a) Compounds of general formula XIII (prepared as described in Scheme 3)were reacted with compounds of general formula by one of the followingmethods 1) for X=mesylate, compounds XIII and VII were reacted in thepresence of a suitable base such as sodium hydride (NaH) and a polarorganic solvent DMF under inert gas conditions at room temperature ii)for X=H, compounds of general formula XIII and VII were reacted usingcustomary Mitsunobu conditions, preferably using Ph₃P (1.4 eq.) and DEAD(1.4 eq.) in organic solvent such as THF under inert gas conditions at atemperature of preferably 70° C. Typical conditions for such reactionsare exemplified in the section C) Side chain introduction conditionsbelow.

b) N—BOC deprotection was performed under customary BOC deprotectionconditions using among the possible acid preferably trifluoro-acteticacid and organic solvent preferably CH₂Cl₂. The reaction was preferablyperformed at room temperature.

c) Amide bond formation was performed using compounds of general formulaXV and an acid chloride of formula R⁴C(O)Cl or carboxylic acid offormula R⁴C(O)OH to prepare an amide of general formula VI; customaryamide bond coupling conditions, as described in Scheme 1, step a) havebeen used. In addition to the methods described in Scheme 1, step a),coupling of carboxylic acids using HOBt/EDC or coupling usingchloroformates or carbamic chlorides were used. The couplings wereperformed in an organic solvent such as preferably DMF or DCM and thefinal compounds were purified by normal or reversed phasechromatography. Typical conditions for amid bond formation reactions areexemplified in the section B) Amide bond formation conditions below.

a) 7-Chloro-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine XVII was preparedfrom 7-chloro-1H-pyrido[3,4-b][1,4]oxazin-2-one XVI by customaryreduction methods, using as reducing agent preferably BH₃*THF and assolvent preferably THF. XVI is available via flow nitration of2-chloro-5-(2-methoxy-2-oxoethoxy)pyridine-1-oxide, followed byreduction and cyclisation.

b) Cross-coupling between XVII and an aryl halogenide of the generalformula R²—X′ where X′=bromo or iodo was performed under customaryBuchwald-Hartwig conditions using a Pd catalyst/ligand combination suchas preferably Pd₂(dba)₃/X-Phos, and a base, such as preferably Cs₂CO₃and an organic solvent such as preferably dioxane. The reaction waspreferably stirred at a temperature of approximately 100° C. and couldbe performed in a microwave reactor. The reaction was preferably carriedout under an inert gas such as nitrogen or argon. Typical conditions forBuchwald-Hartwig cross-coupling reactions are exemplified in the sectionA) Buchwald aminations or hydroxylations below.

c) Hydroxylation of XVIII was performed using aq. KOH and a Pdcatalyst/ligand combination such as preferablyPd₂(dba)₃/tetramethyl-tert-butyl-Xphos and an organic solvent such aspreferably dioxane. The reaction was preferably stirred at a temperatureof approximately 100° C. The reaction was preferably carried out underan inert gas such as nitrogen or argon.

d) Coupling of a compound of general formula XIX with a compound ofgeneral formula VII was performed using a suitable base such as sodiumhydride (NaH, Cs₂CO₃, K₂CO₃) and polar organic solvent such as DMF underinert gas conditions at a temperature of preferably 60-80° C. Typicalconditions for such reactions are exemplified in the section C) Sidechain introduction conditions below.

e) N—BOC deprotection was performed under customary BOC deprotectionconditions using among the possible acid preferably trifluoro-acteticacid and organic solvent preferably CH₂Cl₂. The reaction was preferablyperformed at room temperature.

f) Amide bond formation was performed using compounds of general formulaXXI and an acid chloride of formula R⁴C(O)Cl or carboxylic acid offormula R⁴C(O)OH to prepare an amide of general formula XXII; customaryamide bond coupling conditions, as described in Scheme 1, step a) havebeen used, in addition coupling of carboxylic acids using HOBt/EDC wasapplied. The couplings were performed in an organic solvent such aspreferably DMF or DCM and the final compounds were purified by normal orreversed phase chromatography. Typical conditions for amid bondformation reactions are exemplified in the section B) Amide bondformation conditions below.

a) Hydroxylation of 7-chloro-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazineXVII to give 2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-ol XXIII wasperformed using aq. KOH and a Pd catalyst/ligand combination such aspreferably Pd₂(dba)₃/tetramethyl-tert-butyl-Xphos and an organic solventsuch as preferably dioxane. The reaction was preferably stirred at atemperature of approximately 100° C. The reaction was preferably carriedout under an inert gas such as nitrogen or argon.

b) Coupling of compound XXIII with mesylate VII was effected using asuitable base such as sodium hydride (NaH) and polar organic solventsuch as DMF under inert gas conditions at a temperature of preferably80° C. Typical conditions for such reactions are exemplified in thesection C) Side chain introduction conditions below.

c) Cross-coupling between XXIV and an aryl halogenide of the generalformula R²—X′ where X′=bromo or iodo was performed under customaryBuchwald-Hartwig conditions using a Pd catalyst/ligand combination suchas preferably Pd₂(dba)₃/X-Phos or Pd₂(dba)₃/(rac)-BINAP, and a base,such as preferably Cs₂CO₃ or NaOtBu and an organic solvent such aspreferably dioxane or toluene. The reaction was preferably stirred at atemperature of approximately 100° C. and could be performed in amicrowave reactor. The reaction was preferably carried out under aninert gas such as nitrogen or argon. Typical conditions forBuchwald-Hartwig cross-coupling reactions are exemplified in the sectionA) Buchwald aminations or hydroxylations below.

d) N—BOC deprotection was performed under customary BOC deprotectionconditions using among the possible acid preferably trifluoro-acteticacid and organic solvent preferably CH₂Cl₂. The reaction was preferablyperformed at room temperature

e) Amide bond formation was performed using compounds of general formulaXXI and an acid chloride of formula R⁴C(O)Cl or carboxylic acid offormula R⁴C(O)OH to prepare an amide of general formula XXII; customaryamide bond coupling conditions, as described in Scheme 1, step a) havebeen used or coupling of carboxylic acids using HBTU, HOBt/EDC or HATUwas applied. The couplings were performed in an organic solvent such aspreferably DMF or DCM and the final compounds were purified by normal orreversed phase chromatography. Typical conditions for amid bondformation reactions are exemplified in the section B) Amide bondformation conditions below.

General Chromatography Information

LCMS Method M1 (Rt_(M1))

-   HPLC-column dimensions: 2.1×50 mm-   HPLC-column type: Acquity UPLC HSS T3, 1.8 μm-   HPLC-eluent: A) water+0.05 Vol.-% formic acid+3.75 mM ammonium    acetate B) ACN+0.04 Vol.-% formic acid-   HPLC-gradient: 2-98% B in 1.4 min, 98% B 0.45 min, flow=1.2 ml/min-   HPLC-column temperature: 50° C.

LCMS Method M2 (Rt_(M2))

-   HPLC-column dimensions: 2.1×30 mm-   HPLC-column type: Ascentis Express C18, 2.7 μm-   HPLC-eluent A) water+0.05 Vol.-% formic acid+3.75 mM ammonium    acetate B) ACN+0.04 Vol.-% formic acid-   HPLC-gradient: 2-98% B in 1.4 min, 0.75 min 98% B, flow=1.2 ml/min-   HPLC-column temperature: 50° C.

LCMS Method M3 (Rt_(M3))

-   HPLC-column dimensions: 2.1×30 mm-   HPLC-column type: Ascentis Express C18, 2.7 μm-   HPLC-eluent A) water+0.05 Vol.-% formic acid+3.75 mM ammonium    acetate B) ACN+0.04 Vol.-% formic acid-   HPLC-gradient: 2-98% B in 8.5 min, 1 min 98% B, flow=1.2 ml/min-   HPLC-column temperature: 50° C.

LCMS Method M4 (Rt_(M4))

-   HPLC-column dimensions: 4.6×50 mm-   HPLC-column type: SunFire C18, 5 μm-   HPLC-eluent A) water+0.1 Vol.-% TFA, B) ACN+0.1 Vol.-% TFA-   HPLC-gradient: 5-100% B in 8.0 min B, flow=2 ml/min-   HPLC-column temperature: 40° C.

LCMS Method M5 (Rt_(M5))

-   HPLC-column dimensions: 0.46×25 cm-   HPLC-column type: Chiralcel OJ-H (1189)-   HPLC-eluent EtOH/MeOH 60:40-   HPLC-gradient: isocratic, flow=0.5 ml/min-   Detector: UV 220 nm

LCMS Method M6 (Rt_(M6))

-   HPLC-column dimensions: 2.1×30 mm-   HPLC-column type: Ascentis Express C18, 2.7 μm-   HPLC-eluent A) water+0.05% TFA, B) ACN+0.04% TFA-   HPLC-gradient: 2-98% B in 1.4 min, 0.75 min 98% B, flow=1.2 ml/min-   HPLC-column temperature: 50° C.

LCMS Method M7 (Rt_(M7))

-   HPLC-column dimensions: 2.1×30 mm-   HPLC-column type: Ascentis Express C18, 2.7 μm-   HPLC-eluent A) water+0.05% TFA, B) ACN+0.04% TFA-   HPLC-gradient: 10-95% B in 3.0 min, 1 min 95% B, flow=1.2 ml/min-   HPLC-column temperature: 50° C.

LCMS Method M8 (Rt_(M8))

-   HPLC-column dimensions: 2.1×30 mm-   HPLC-column type: Ascentis Express C18 2.7 μm-   HPLC-eluent: A) water+0.05% formic acid+3.75 mM ammonium acetate, B)    acetonitrile+0.04% formic acid-   HPLC-gradient: 10-95% B in 3.0 min, flow=1.2 ml/min

LCMS Method M9 (Rt_(M9))

-   HPLC-column dimensions: 2.1×30 mm-   HPLC-column type: Ascentis Express C18 2.7 μm-   HPLC-eluent: A) water+0.05% formic acid+3.75 mM ammonium acetate, B)    acetonitrile+0.04% formic acid-   HPLC-gradient: 10% B from 0.0 to 0.5 min then from 0.5 min to 3.0    min gradient 10-95% B, flow=1.2 ml/min

LCMS Method M10 (Rt_(M10))

-   HPLC-column dimensions: 2.1×50 mm-   HPLC-column type: Acquity UPLC BEH C18 1.7 μm-   HPLC-eluent: A) water+0.1 Vol.-% formic acid, B) acetonitrile-   HPLC-gradient: 20-25% B in 1.00 min, then 25-95% B in 3.20 min, then    95-100% B in 0.10 min, then 100% for 0.20 min, flow=0.7 ml/min

LCMS Method M11 (Rt_(M11))

-   HPLC-column dimensions: 2.1×50 mm-   HPLC-column type: Acquity UPLC BEH C18 1.7 μm-   HPLC-eluent: A) water+0.1 Vol.-% formic acid, B) acetonitrile-   HPLC-gradient: 5-10% B in 1.00 min, then 10-90% B in 3.00 min, then    90-100% B in 0.10 min, then 100% for 0.40 min, flow=0.7 ml/min

LCMS Method M12 (Rt_(M12))

-   HPLC-column dimensions: 2.1×30 mm-   HPLC-column type: Ascentis Express C18 2.7 μm-   HPLC-eluent: A) water+0.1 Vol.-% TFA, B) acetonitrile-   HPLC-gradient: 10-95% B over 1.7 min and 1.2 mL/min as solvent flow    and then 95 5 B over 0.7 min, flow=1.4 mL/min.

LCMS Method M13 (Rt_(M13))

-   HPLC-column dimensions: 2.1×30 mm-   HPLC-column type: Ascentis Express C18 2.7 μm-   HPLC-eluent: A) water+0.05% formic acid+3.75 mM ammonium acetate, B)    acetonitrile+0.04% formic acid-   HPLC-gradient: 10-95% B in 3.7 min, flow=1.2 ml/min

LCMS Method M14 (Rt_(M14))

-   HPLC-column dimensions: 2.1×30 mm-   HPLC-column type: Ascentis Express C18, 2.7 μm-   HPLC-eluent A) water+0.05% formic acid+3.75 mM ammonium acetate, B)    acetonitrile+0.04% formic acid-   HPLC-gradient: 10-95% B in 1.5 min, 1 min 95% B, flow=1.2 ml/min

LCMS Method M15 (Rt_(M15))

-   HPLC-column dimensions: 0.46×25 cm-   HPLC-column type: Chiralcel OD-H (1194)-   HPLC-eluent Hexan/EtOH 50:50+0.05% DEA-   HPLC-gradient: isocratic, flow=0.5 ml/min-   Detector: UV 220 nm

LCMS Method M16 (Rt_(M16))

-   HPLC-column dimensions: 2.1×50 mm-   HPLC-column type: Acquity UPLC HSS T3, 1.8 μm-   HPLC-eluent: A) water+0.05 Vol.-% formic acid+3.75 mM ammonium    acetate B) ACN+0.04 Vol.-% formic acid-   HPLC-gradient: 5-98% B in 1.4 min, 98% B 0.4 min, flow=1.0 ml/min-   HPLC-column temperature: 60° C.

X-Ray Powder Diffraction

Instrumentation:

Method X1

Instrument Bruker D8 GADDS Discover

Irradiation CuKα (40 kV, 40 mA)

Detector HI-STAR Area detector

Scan range 6°-39° (2 theta value)

Melting Point Determination:

Melting point was determined by Differential Scanning calorimetry (DSC).DSC was as recorded on a TA Instruments DSC Q2000 using a heating rateof 10° C./min. A sample of 0.6 mg was weighed into standard aluminiumpan (pan+lid, TA 900786.901, 900779.901). The instrument was operatedusing the Thermal Advantage Q-Series software V.2.6.0.367 and theThermal Advantage software V4.6.9. Thermal events were characterizedusing Universal Analysis V4.3A Build 4.3.0.6. The samples was measuredagainst sample pan without pin hole. The sample was treated according tothe protocol below:

Step 1: EQUILIBRATE AT 0° C.

Step 2: Ramp 10° C./min to 300° C.

Preparation of Examples

Where it is stated that compounds were prepared in the manner describedfor an earlier example, the skilled person will appreciate that reactiontimes, number of equivalents of reagents and reaction temperatures maybe modified for each specific reaction, and that it may nevertheless benecessary or desirable to employ different work-up or purificationconditions.

Example A1(S)-(3-((4-(6-methoxy-5-methylpyridin-3-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)oxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone(according to Scheme 1) a1)(R)-(3-hydroxypyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone

A stirred solution of tetrahydro-2H-pyran-4-carboxylic acid (CASregistry 5337-03-1) (0.200 g, 1.537 mmol) and DMF (0.012 ml, 0.154 mmol)in DCM (3 ml) was treated with oxalyl chloride (0.202 ml, 2.305 mmol) at3° C. After 1 h at 3° C., the reaction mixture was concentrated underreduced pressure. The residue was then dissolved in DCM (2 ml), andadded to a stirred solution of (R)-pyrrolidin-3-ol hydrochloride (CASregistry 104706-47-0) (0.190 g, 1.537 mmol), Et₃N (0.535 ml, 3.84 mmol)in DCM (3 ml) at 3° C. After 1 h at 3° C., the reaction mixture wasconcentrated under reduced pressure. The residue was treated with EtOAc(10 ml) and filtered. The residue was washed with EtOAc, and thecombined filtrates were concentrated under reduced pressure. The crudeproduct was purified by flash chromatography on silica gel (DCM/methanolgradient) to provide the title compound as a white solid.

ESIMS: 200 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 4.61-4.50 (m, 1H), 4.10-4.02 (m, 2H),3.77-3.40 (m, 6H), 2.70-2.53 (m, 1H), 2.20-1.85 (m, 4H), 1.75-1.69 (m,3H).

alternative method a2: instead of preparing the acid chloride in situ, acommercially available acid chloride like propanoyl chloride (CASregistry 79-03-8) was used.

b1) (R)-1-(tetrahydro-2H-pyran-4-carbonyl)pyrrolidin-3-ylmethanesulfonate

A stirred solution of(R)-(3-hydroxypyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone (0.245g, 1.230 mmol) in DCM (10 ml) was treated with Et₃N (0.343 ml, 2.459mmol) and methanesulfonyl chloride (0.192 ml, 2.459 mmol) at 0° C. After1 h at 0° C., water (20 ml) was added. The organic layer was washed witha saturated NaCl solution (20 ml), dried with MgSO₄, filtered andconcentrated under reduced pressure. The crude product was purified bytrituration with diethyl ether to provide the title compound as a whitesolid.

ESIMS: 278 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 5.40-5.29 (m, 1H), 4.10-4.02 (m, 2H),3.94-3.87 (m, 1H), 3.82-3.56 (m. 3H), 3.52-3.41 (m, 2H), 3.11-3.04 (m,3H), 2.70-2.10 (m, 3H), 2.02-2.87 (m, 2H), 1.72-1.57 (m, 2H).

c1)(S)-(3-((3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)oxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone

A stirred solution of 3,4-dihydro-2H-benzoxazin-6-ol (CAS registry26021-57-8) (0.140 g, 0.926 mmol) in DMF (3 ml) was treated with sodiumhydride (60% in mineral oil, 0.445 g, 1.111 mmol) at rt. After 10 min atrt, (R)-1-(tetrahydro-2H-pyran-4-carbonyl)pyrrolidin-3-ylmethanesulfonate (0.283 g, 1.019 mmol) was added. The vial was cappedand heated to 50° C. for 3 h. After this time, the reaction mixture wasconcentrated under reduced pressure. The residue was dissolved in EtOAc(50 ml), and water (50 ml) was added. The organic layer was washed witha saturated NaCl solution (20 ml), dried with MgSO₄, filtered andconcentrated under reduced pressure. The crude product was purified byflash chromatography on silica gel (DCM/methanol gradient) to providethe title compound as a grey amorphous solid.

HPLC Rt_(M10)=2.07 min; ESIMS: 333 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 6.55-6.50 (m, 1H), 6.15-6.11 (m, 1H),6.07-6.00 (m, 1H), 5.77 (br s, 1H), 4.88-4.74 (m, 1H), 4.06-4.01 (m,2H), 3.90-3.22 (m, 10H), 2.75-2.58 (m, 1H), 2.15-1.95 (m, 2H), 1.65-1.45(m, 4H).

d1)(S)-(3-((4-(6-methoxy-5-methylpyridin-3-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)oxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone

A stirred solution of(S)-(3-((3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)oxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone(0.050 g, 0.150 mmol) in toluene (1 ml) was treated with5-bromo-2-methoxy-3-methylpyridine (CAS registry 760207-87-2) (0.030 g,0.150 mmol), NaOtBu (0.022 g, 0.226 mmol),2-(dicyclohexylphosphino)biphenyl (CAS registry 247940-06-3) andPd₂(dba)₃ (0.004 g, 0.005 mmol) at rt under argon. The reaction vial wascapped and heated to 110° C. in a microwave reactor for 3 h. After thistime, the reaction mixture was concentrated under reduced pressure. Thecrude product was purified by flash chromatography on silica gel(cyclohexane/EtOAc gradient) to provide the title compound as anoff-white solid.

HPLC Rt_(M10)=2.85 min; ESIMS: 454 [(M+H)⁺].

¹H NMR (400 MHz, CD₃OD): δ 7.90-7.85 (m, 1H), 7.49-7.44 (m, 1H),6.76-6.69 (m, 1H), 6.32-6.24 (m, 1H), 6.07-6.02 (m, 1H), 4.86-4.73 (m,1H), 4.29-4.23 (m, 2H), 4.02-3.92 (m, 5H), 3.80-3.40 (m, 8H), 2.85-2.60(m, 1H), 2.25-1.91 (m, 5H), 1.85-1.50 (m, 4H).

alternative method d2: 2-(dicyclohexylphosphino)biphenyl (CAS registry247940-06-3) was replaced with2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (CAS registry564483-18-7)

alternative method d3: 2-(dicyclohexylphosphino)biphenyl (CAS registry247940-06-3) and Pd₂(dba)₃ was replaced withbis(tri-t-butylphosphine)palladium (CAS registry 53199-31-8)

Examples A2 to A43

The compounds listed in Table 1 were prepared by a procedure analogousto that used in Example A1.

TABLE 1 HPLC Rt MS [min] [m/z; Example Compound (method) (M + 1)⁺] A2

2.50 (M10) 517 A3

1.10 (M12) 560 A4

1.78 (M10) 469 A5

1.03 (M10) 425 A6

1.21 (M12) 479 A7

2.44 (M10) 435 A8

1.46 (M8) 432 A9

2.80 (M10) 409 A10

2.74 (M11) 452 A11

2.86 (M10) 402 A12

3.00 (M10) 418 A13

2.93 (M10) 398 A14

1.98 (M10) 399 A15

2.76 (M10) 490 A16

2.56 (M10) 461 A17

2.67 (M10) 437 A18

1.60 (M9) 546 A19

1.32 (M9) 424 A20

1.47 (M9) 454 A21

1.61 (M9) 478 A22

1.43 (M9) 440 A23

1.57 (M9) 493 A24

2.04 (M9) 514 A25

1.83 (M9) 532 A26

1.83 (M9) 503 A27

2.22 (M9) 545 A28

2.08 (M9) 516 A29

2.82 (M9) 506 A30

2.69 (M9) 477 A31

2.72 (M9) 492 A32

2.49 (M9) 379 A33

1.54 (M9) 440 A34

1.76 (M9) 444 A35

1.24 (M6) 413 A36

1.02 (M6) 404 A37

1.03 (M6) 432 A38

1.24 (M6) 422 A39

1.13 (M6) 461 A40

1.33 (M6) 392 A41

1.44 (M6) 451 A42

1.82 (M6) 478 A43

1.44 (M6) 488

Example B1{(S)-3-[4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone(according to Scheme 2)

a)(S)-3-(3,4-Dihydro-2H-benzo[1,4]oxazin-6-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

A solution of 3,4-dihydro-2H-benzo[1,4]oxazin-6-ol (CAS registry26021-57-8) (4.0 g, 26.5 mmol) in DMF (150 ml) was treated with NaH(2.117 g, 52.9 mmol) for 20 min at 20° C.(R)-3-Methanesulfonyloxy-pyrrolidine-1-carboxylic acid tert-butyl ester(CAS registry 127423-61-4) (9.13 g, 34.4 mmol) was added. After stirringfor 22 h at rt the reaction mixture was concentrated to dryness, thentaken up with EtOAc, filtered through hyflo and the filtrate was washedwith sat. aq. Na₂CO₃ solution. Combined organic layers were washed withbrine, dried over Na₂SO₄, filtered and evaporated. The crude product waspurified by flash chromatography on silica gel (cyclohexane/isopropanol100:0 to 85:15 in 40 min) to provide the title compound as a yellow oil.

HPLC Rt_(M8)=1.84 min; ESIMS: 321 [(M+H)⁺].

¹H NMR (400 MHz, DMSO): 6.52 (d, 1H), 6.12 (d, 1H), 6.02 (m, 1H), 5.76(m, 1H), 4.75 (br s, 1H), 4.01-40.5 (m, 2H), 3.27-3.50 (m, 4H),3.22-3.26 (m, 2H), 1.95-2.08 (m, 2H), 1.39 (m, 9H).

b)(S)-3-[4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

A mixture of(S)-3-(3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (2.12 g, 6.62 mmol),5-bromo-2-methanesulfonyl-3-methyl-pyridine (Intermediate IA1) (2.091 g,7.94 mmol), NaOtBu (1.272 g, 13.23 mmol), XPhos ligand (0.158 g, 0.331mmol) and Pd₂(dba)₃ (0.303 g, 0.331 mmol) in dioxane (3.5 ml) wasdegassed and stirred for 12 h at 110° C. Sat. aq. NaHCO₃ solution wasadded and the reaction mixture was extracted with EtOAc. Combinedorganic layers were washed with brine, dried over Na₂SO₄, filtered andevaporated. The crude product was purified by flash chromatography onsilica gel (cyclohexane/EtOAc 100:0 to 50:50) to provide the titlecompound.

HPLC Rt_(M14)=1.25 min; ESIMS: 490 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.33 (d, 1H), 7.43 (d, 1H), 6.86 (d, 1H),6.59 (d, 1H), 6.47 (m, 1H), 4.69-4.73 (m, 12H), 4.23-4.28 (m, 2H),3.73-3.78 (m, 2H), 3.41-3.58 (m, 4H), 3.34 (s, 3H), 2.69 (s, 3H),1.96-2.17 (m, 2H), 1.46 (s, 9H).

c)4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-6-((S)-pyrrolidin-3-yloxy)-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of(S)-3-[4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (1.5 g, 3.06 mmol) and TFA (0.236 ml, 3.06 mmol)in DCM (15 ml) was stirred for 1 h at rt. The reaction mixture wascooled down to 0° C., sat. Na₂CO₃ solution was added and the reactionmixture was extracted with DCM. Combined organic layers were dried overNa₂SO₄, filtered and evaporated to provide the title compound.

HPLC Rt_(M2)=0.66 min; ESIMS: 390 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.33 (d, 1H), 7.43 (d, 1H), 6.86 (d, 1H),6.58 (d, 1H), 6.47 (m, 1H), 4.68 (m, 1H), 4.22-4.27 (m, 2H), 3.73-3.78(m, 2H), 3.33 (s, 3H), 3.12-3.22 (m, 2H), 2.86-3.04 (m, 2H), 2.68 (s,3H), 1.88-2.08 (m, 2H).

d){(S)-3-[4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

A mixture of4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-6-((S)-pyridin-3-yloxy)-3,4-dihydro-2H-benzo[1,4]oxazine(0.085 g, 0.218 mmol), tetrahydro-2H-pyran-4-carbonyl chloride (CASregistry 40191-32-0) (0.049 mg, 0.327 mmol) and Et₃N (0.046 ml, 0.327mmol) in DCM (4 ml) was stirred at rt for 15 min. The reaction mixturewas concentrated to dryness. The crude product was purified by prep.RP-HPLC (column SunFire C18, H₂O+0.1% TFA/ACN+0.1% TFA 90:10 to 30:70 in12 min) to provide the title compound as a white solid.

HPLC Rt_(M7)=1.62 min; ESIMS: 502 [(M+H)⁺].

¹H NMR (400 MHz, DMSO): δ 8.38-8.42 (m, 1H), 7.72 (m, 1H), 6.84 (d, 1H),6.67 (m, 1H), 6.50-6.57 (m, 1H), 4.82-4.94 (m, 1H), 4.20 (m, 2H), 3.31(s, 3H), 3.28-3.88 (m, 10H), 2.59-2.73 (m, 1H), 2.56 (s, 3H), 1.95-2.13(m, 2H), 1.44-1.62 (m, 4H).

Examples B2 to B122

The compounds listed in Table 2 were prepared by a procedure analogousto that used in Example B1.

TABLE 2 HPLC Rt MS Compound/ [min] [m/z; Example Reaction Conditions(method) (M + 1)⁺] B2

 2.00 (M8) 468 B3

 1.53 (M8) 379 B4

 1.58 (M9) 414 B5

 2.61 (M10) 460 B6

 1.50 (M9) 560 B7

 1.37 (M8) 486 B8

 1.27 (M8) 498 B9

 1.34 (M8) 518 B10

 1.68 (M8) 522 B11

 1.67 (M8) 506 B12

 1.80 (M8) 522, 524 B13

 1.74 (M8) 488 B14

 1.95 (M8) 556 B15

 1.89 (M7) 440 B16

 1.89 (M7) 440 B17

 1.88 (M8) 516 B18

 1.86 (M7) 458 B19

 1.04 (M2) 516 B20

 1.70 (M8) 450 B21

 1.87 (M8) 450 B22

 0.85 (M6) 495 B23

 0.92 (M6) 496 B24

 0.95 (M6) 485 B25

 0.94 (M6) 485 B26

 1.01 (M6) 530 B27

 1.62 (M2) 488 B28

 1.62 (M2) 488 B29

 1.53 (M6) 564 B30

 1.50 (M9) 464 B31

 0.94 (M6) 488 B32

 0.92 (M6) 462 B33

 1.01 (M6) 460 B34

 1.05 (M6) 494 B35

 0.87 (M2) 536 B36

 0.88 (M2) 504 B37

 0.88 (M2) 476 B38

 0.93 (M2) 470 B39

 0.91 (M2) 472 B40

 0.92 (M2) 456 B41

 0.91 (M2) 504 B42

 0.91 (M2) 502 B43

 0.96 (M2) 470 B44

 0.90 (M2) 430 B45

 0.89 (M2) 492 B46

 0.92 (M2) 444 B47

 0.90 (M2) 518 B48

 0.85 (M2) 466 B49

 1.06 (M2) 500 B50

 0.86 (M2) 516 B51

 0.89 (M2) 516 B52

 0.88 (M2) 15.13 (CD10) 488 B53

 0.88 (M2) 18.70 (CD10) 488 B54

 3.21 (M3) 474, 476 B55

 3.30 (M3) 418, 420 B56

 3.02 (M3) 522, 524 B57

 0.88 (M2) 434, 436 B58

 3.36 (M3) 458, 460 B59

 3.47 (M3) 402, 404 B60

 3.17 (M3) 506, 508 B61

 0.91 (M2) 418, 420 B62

 2.59 (M3) 469 B63

 2.85 (M3) 482, 483 B64

 2.71 (M3) 453 B65

 2.97 (M3) 466, 468 B66

 0.88 (M2) 484 B67

 0.86 (M2) 509 B68

 0.77 (M2) 509 B69

 0.93 (M1) 556 B70

 0.91 (M1) 538 B71

 0.88 (M1) 520 B72

 1.07 (M1) 490 B73

 1.01 (M1) 538 B74

 0.81 (M1) 534 B75

 0.88 (M1) 524 B76

 0.88 (M1) 524 B77

 0.85 (M1) 517 B78

 0.86 (M1) 531 B79

 0.98 (M1) 522 B80

 0.94 (M1) 458 B81

 0.95 (M1) 538 B82

 0.99 (M1) 496 B83

 0.96 (M1) 494 B84

 0.94 (M1) 482 B85

 1.08 (M1) 494 B86

 0.80 (M1) 506 B87

 1.57 (M7) 502 B88

 1.45 (M8) 450 B89

 1.52 (M8) 450 B90

 1.69 (M8) 495 B91

 1.53 (M8) 463 B92

 1.54 (M8) 463 B93

 1.71 (M8) 502 B94

 1.88 (M8) 440 B95

 1.59 (M8) 477 B96

 1.50 (M7) 446 B97

 1.53 (M8) 446 B98

 1.83 (M7) 444 B99

 1.77 (M7) 418 B100

 1.87 (M7) 441 B101

 1.81 (M7) 492 B102

 1.48 (M7) 453 B103

 1.80 (M7) 441 B104

 1.75 (M7) 452 B105

 1.84 (M8) 508, 510 B106

 1.78 (M8) 496, 498 B107

 1.67 (M8) 434, 436 B108

 1.84 (M8) 457, 459 B109

 1.88 (M8) 429, 431 B110

 1.90 (M8) 482 B111

 1.70 (M8) 466 B112

 1.77 (M8) 480 B113

 1.57 (M8) 418 B114

 1.84 (M8) 432 B115

 1.83 (M8) 459 B116

 1.85 (M8) 413 B117

 3.13 (M3) 472 B118

 3.40 (M3) 472 B119

 0.97 (M1) 538 B120

 1.03 (M1) 490 B121

 1.42 (M8) 430 B122

 1.66 (M8) 481

Example C12-Methoxy-5-{6-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile(according to Scheme 3)

a) 6-(tert-Butyl-dimethyl-silanyloxy)-3,4-dihydro-2H-benzo[1,4]oxazine

Under argon, NaH (2.96 g, 74.1 mmol) was portionwise added to a solutionof 3,4-dihydro-2H-benzo[1,4]oxazin-6-ol (CAS registry 26021-57-8) (5.60g, 37.0 mmol) in THF (200 ml). After stirring at rt for 20 min, TBDMSCl(CAS registry 18162-48-6) (7.26 g, 48.2 mmol) was slowly added andstirring was continued for 1 h. The reaction mixture was diluted withEt₂O, washed with a sat. aq. NaHCO₃ soln. and brine. The organic phasewas dried over MgSO₄, concentrated and the title compound was obtainedafter flash chromatography on silica gel (cyclohexane/EtOAc 100:0 to60:40 over 15 min) as a yellow oil (9.20 g, 94% yield).

HPLC Rt_(M10)=3.65 min; ESIMS: 266 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 6.46 (d, 1H), 6.08 (d, 1H), 5.91 (m, 1H),5.71 (br s, 1H), 3.91-4.12 (m, 2H), 3.12-3.28 (m, 2H), 0.87-1.01 (s,9H), 0.03-0.21 (s, 3H).

b)5-[6-(tert-Butyl-dimethyl-silanyloxy)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-2-methoxynicotinonitrile

Under argon, XPhos (CAS registry 564483-18-7) (0.79 g, 1.7 mmol) andPd₂(dba)₃ (CAS registry 51364-51-3) (1.52 g, 1.7 mmol) were added to asuspension of6-(tert-butyl-dimethyl-silanyloxy)-3,4-dihydro-2H-benzo[1,4]oxazine(9.00 g, 33.2 mmol), 5-bromo-2-methoxy-nicotinonitrile (CAS registry941294-54-8) (7.79 g, 36.6 mmol), NaOtBu (4.79 g, 49.8 mmol) in toluene(270 ml). The reaction mixture was stirred at 110° C. for 1 h and wasconcentrated to afford a brown solid which was washed with a mixture ofDCM/MeOH (8:2) and filtered off. The filtrate was concentrated, theobtained residue was dissolved in DCM/MeOH (8:2), filtered over hyflo,the filtrate was concentrated and triturated with MeOH to afford thetitle compound as yellow solid (10.14 g, 77% yield).

HPLC Rt_(M11)=3.89 min; ESIMS: 398 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.35-8.51 (m, 1H), 8.16-8.31 (m, 1H),6.60-6.79 (m, 1H), 6.15-6.32 (m, 1H), 5.92-6.09 (m, 1H), 4.00 (s, 3H),3.51-3.74 (m, 2H), 0.87 (s, 9H), 0.07 (s, 6H).

c)5-(6-hydroxy-2,3-dihydro-benzo[1,4]oxazin-4-yl)-2-methoxy-nicotinonitrile

TBAF (1M in THF) (37.7 ml, 37.7 mmol) was added to a solution of5-[6-(tert-butyl-dimethyl-silanyloxy)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-2-methoxy-nicotinonitrile(10 g, 25.2 mmol) dissolved in THF (200 ml). The solution was stirred atrt for 30 min, diluted with EtOAc, washed with sat. aq. NaHCO₃ soln. andbrine. The aqueous layers were back extracted with EtOAc, concentrationof the organic phases after drying over MgSO₄ afforded a brown residuewhich was dissolved in DCM/MeOH (1:1) and filtered over hyflo.Concentration and trituration with Et₂O of the filtrate afforded thetitle compound as brown solid (6.63 g, 93% yield).

HPLC Rt_(M10)=2.56 min; ESIMS: 284 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.70 (br. s, 1H), 8.44 (d, 1H), 8.28 (d,1H), 6.62 (d, 1H), 6.12 (m, 1H), 6.01 (d, 1H), 4.11-4.32 (m, 2H), 4.01(s, 3H), 3.54-3.68 (m, 2H).

d)2-Methoxy-5-{6-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile

Under argon, NaH (31 mg, 0.78 mmol) was added to a solution of5-(6-hydroxy-2,3-dihydro-benzo[1,4]oxazin-4-yl)-2-methoxy-nicotinonitrile(100 mg, 0.35 mmol) in DMF (2 ml) and stirred at rt for 5 min.Methanesulfonic acid (R)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-ylester (intermediate IC1) (98.0 mg, 0.35 mmol) was added and the reactionmixture was stirred at 50° C. for 4 h. After cooling, NaH (0.5 eq., 8.47mg, 0.21 mmol) was added, the reaction mixture was stirred at rt for 5min and methanesulfonic acid(R)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yl ester (intermediateIC1) (49.0 mg, 0.18 mmol) was added. The reaction mixture was stirred at50° C. for 1 h. Concentration and purification by prep. RP-HPLC (SunfirePrepC18 OBD 30×100 mm, 5 μm; solvent A: H₂O+0.1 Vol.-% TFA; solvent B:CH₃CN+0.1 Vol.-% TFA) afforded, after basification of the combinedfractions and extraction with EtOAc, the title compound as a yellowsolid (72 mg, 43% yield).

HPLC Rt_(M10)=2.72 min; ESIMS: 465 [(M+H)⁺].

¹H NMR (400 MHz, CD₃OD): δ 8.36 (d, 1H), 8.06 (t, 1H), 6.78 (m, 1H),6.37 (m, 1H), 6.17 (m, 1H), 4.81 (br s, 1H), 4.17-4.37 (m, 2H), 4.08 (s,3H), 3.90-4.03 (m, 2H), 3.56-3.81 (m, 5H), 3.39-3.54 (m, 3H), 2.59-2.89(m, 1H), 1.87-2.29 (m, 2H), 1.48-1.87 (m, 4H).

Examples C2 to C26

The compounds listed in Table 3 were prepared by a procedure analogousto that used in Example C1.

TABLE 3 UPLC Rt [min] MS [m/z; Example Compound (method) (M + 1)⁺] C2

 2.71 (M10) 465 C3

 2.71 (M10) 384 C4

 2.71 (M10) 384 C5

 2.83 (M10)  16.981 (M5) 479 C6

 2.83 (M10)  19.957 (M5) 479 C7

 4.13 (M4) 449 C8

 4.11 (M4) 461 C9

 4.17 (M4) 448 C10

 3.60 (M4) 447 C11

 3.73 (M4) 473 C12

 4.02 (M4) 463 C13

 3.72 (M4) 459 C14

 4.02 (M4) 464 C15

 4.03 (M3) 459 C16

 3.27 (M3) 458 C17

 3.20 (M3) 458 C18

 3.91 (M3) 475 C19

 3.44 (M3) 464 C20

 4.27 (M3) 476 C21

 4.33 (M3) 505 C22

 3.87 (M3) 498 C23

 3.47 (M3) 464 C24

 2.68 (M10) 22.58 (CD7) 467 C25

 2.68 (M10) 33.80 (CD7) 467 C26

 1.57 (M9) 465

Example D1(S)-2-methoxy-5-(6-((1-(1-methyl-1H-imidazole-4-carbonyl)pyrrolidin-3-yl)oxy)-2H-benzo[b][1,4]oxazin-4(3H)-yl)nicotinonitrile(according to Scheme 4) a1)6-((tert-butyldimethylsilyl)oxy)-3,4-dihydro-2H-benzo[b][1,4]oxazine

A stirred solution of 3,4-dihydro-2H-1,4-benzoxazin-6-ol (CAS registry226021-57-8) (6.00 g, 39.70 mmol) in THF (200 ml) was treated withsodium hydride (60% in mineral oil, 3.18 g, 79.00 mmol) at rt. After 20min at rt, TBDMSCl (7.78 g, 51.6 mmol) was added, and the reactionmixture was stirred at rt for 1.5 h. After that time, diethyl ether (500ml) and a sat. aq. NaHCO₃ soln. (100 ml) were added. The aq. layer wasextracted with diethyl ether, and the combined organic extracts weredried with MgSO₄, filtered and concentrated under reduced pressure. Thecrude product was purified by flash chromatography on silica gel(cyclohexane/EtOAc gradient) to provide the title compound as a yellowoil.

HPLC Rt_(M11)=3.37 min; ESIMS: 266 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 6.48-6.44 (m, 1H), 6.09-6.05 (m, 1H),5.94-5.89 (m, 1H), 5.76-5.70 (m, 1H), 4.06-4.00 (m, 2H), 3.25-3.19 (m,2H), 0.92 (s, 9H), 0.12 (s, 6H).

b1)5-(6-((tert-butyldimethylsilyl)oxy)-2H-benzo[b][1,4]oxazin-4(3H)-yl)-2-methoxynicotinonitrile

A stirred solution of6-((tert-butyldimethylsilyl)oxy)-3,4-dihydro-2H-benzo[b][1,4]oxazine(8.88 g, 32.80 mmol) in toluene (270 ml) was treated with5-bromo-2-methoxynicotinonitrile (CAS registry 941294-54-8) (7.68 g,36.10 mmol), NaOtBu (4.87 g, 49.2 mmol),2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (CAS registry564483-18-7) (0.806 g, 1.64 mmol), and Pd₂dba₃ (1.501 g, 1.64 mmol) atrt under argon. The reaction mixture was heated to 110° C. for 1.5 h.After that time, the reaction mixture was concentrated under reducedpressure. The residue was dissolved in DCM (200 ml), filtered through apad of celite, and concentrated under reduced pressure. The residue wasdissolved in MeOH, and sonicated several times to give a yellow/orangeprecipitate. The residue was filtered, washed with methanol, and driedunder vacuum to provide the title compound as a yellow solid.

HPLC Rt_(M11)=3.90 min; ESIMS: 398 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.45-8.42 (m, 1H), 8.28-8.24 (m, 1H),6.72-6.68 (m, 1H), 6.24-6.19 (m, 1H), 6.06-6.03 (m, 1H), 4.24-4.18 (m,2H), 4.00 (s, 3H), 3.66-3.61 (m, 2H), 0.87 (s, 9H), 0.07 (s, 6H).

alternative method b2:dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (CAS registry564483-18-7) and Pd₂(dba)₃ were replaced withbis(tri-t-butylphosphine)palladium (CAS registry 53199-31-8)

c1)5-(6-hydroxy-2H-benzo[b][1,4]oxazin-4(3H)-yl)-2-methoxynicotinonitrile

A stirred solution of5-(6-((tert-butyldimethylsilyl)oxy)-2H-benzo[b][1,4]oxazin-4(3H)-yl)-2-methoxynicotinonitrile(10.85 g, 27.30 mmol) in THF (220 ml) was treated with TBAF (1.0 M inTHF, 40.9 ml, 40.90 mmol) at rt. After 40 min at rt, EtOAc (300 ml) anda sat. aq. NaHCO₃ soln. (200 ml) were added. The organic extracts weredried with MgSO₄, filtered and concentrated under reduced pressure. Thecrude product was purified by trituration with diethyl ether to providethe title compound as a pale brown solid.

HPLC Rt_(M11)=2.00 min; ESIMS: 284 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.71 (s, 1H), 8.44 (d, 1H), 8.29 (d, 1H),6.61 (d, 1H), 6.12 (dd, 1H), 6.01 (d, 1H), 4.21-4.16 (m, 2H), 4.01 (s,3H), 3.64-3.59 (m, 2H).

d1) (S)-tert-butyl3-((4-(5-cyano-6-methoxypyridin-3-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)oxy)pyrrolidine-1-carboxylate

A stirred solution of5-(6-hydroxy-2H-benzo[b][1,4]oxazin-4(3H)-yl)-2-methoxynicotinonitrile(3.70 g, 13.06 mmol) in DMF (60 ml) was treated with sodium hydride (60%in mineral oil, 1.31 g, 32.70 mmol) at rt. The reaction mixture wasstirred at rt for 15 min. After that time,(R)-1-Boc-3-methanesulfonyloxypyrrolidine (CAS registry 141699-57-2)(5.36 g, 19.59 mmol) was added, and the reaction mixture was stirred at50° C. for 3 h. After that time, the reaction mixture was concentratedunder reduced pressure. The crude product was purified by flashchromatography on silica gel (cyclohexane/acetone gradient) to providethe title compound as a yellow solid.

HPLC Rt_(M11)=3.13 min; ESIMS: 453 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.31 (d, 1H), 7.81 (d, 1H), 6.82 (d, 1H),6.32 (dd, 1H), 6.14 (d, 1H), 4.74-4.68 (m, 1H), 4.32-4.28 (m, 2H), 4.09(s, 3H), 3.67-3.62 (m, 2H), 3.59-3.39 (m, 4H), 2.17-1.92 (m, 2H), 1.47(s, 9H).

alternative method d2: the mesylated alcohol, sodium hydride and DMFwere replaced with the corresponding hydroxy-isoxazolidine, DEAD and THFusing Mitsunobu conditions described in method CC4

e1)(S)-2-methoxy-5-(6-(pyrrolidin-3-yloxy)-2H-benzo[b][1,4]oxazin-4(3H)-yl)nicotinonitrile

A stirred solution of (S)-tert-butyl3-((4-(5-cyano-6-methoxypyridin-3-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)oxy)pyrrolidine-1-carboxylate(4.35 g, 9.32 mmol) in DCM (160 ml) was treated with TFA (35.9 ml, 466mmol) at rt. The reaction mixture was stirred at rt for 2 h. After thattime, the reaction mixture was concentrated under reduced pressure. Theresidue was dissolved in DCM (500 ml), and a saturated aqueous NaHCO₃solution (500 ml) was added. The organic extracts were washed with asaturated aqueous NaCl solution (50 ml), dried with MgSO₄, filtered andconcentrated under reduced pressure. The crude product (title compound,yellow solid) was used in the next step without further purification.

HPLC Rt_(M10)=2.06 min; ESIMS: 353 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.31 (d, 1H), 7.82 (d, 1H), 6.82 (d, 1H),6.32 (dd, 1H), 6.12 (d, 1H), 4.71-4.65 (m, 1H), 4.32-4.27 (m, 2H), 4.09(s, 3H), 3.67-3.62 (m, 2H), 3.22-2.90 (m, 4H), 2.08-1.88 (m, 2H).

f1)(S)-2-methoxy-5-(6-((1-(1-methyl-1H-imidazole-4-carbonyl)pyrrolidin-3-yl)oxy)-2H-benzo[b][1,4]oxazin-4(3H)-yl)nicotinonitrile

A stirred solution of 1-methyl-1H-imidazole-4-carboxylic acid (CASregistry 41716-18-1) (0.578 g, 4.45 mmol) in DMF (40 ml) was treatedwith HOBT (0.695 g, 4.45 mmol), EDC (0.870 g, 4.45 mmol) and Et₃N (1.24ml, 8.90 mmol) at rt. The reaction mixture was stirred at rt for 15 min.After that time,(S)-2-methoxy-5-(6-(pyrrolidin-3-yloxy)-2H-benzo[b][1,4]oxazin-4(3H)-yl)nicotinonitrile(1.10 g, 2.97 mmol) was added, and the reaction mixture was stirred for3 h 15 min at rt. After that time, the reaction mixture was concentratedunder reduced pressure. The residue was dissolved in DCM (200 ml), and asat. aq. NaHCO₃ soln. (200 ml) was added. The organic extracts weredried with MgSO₄, filtered and concentrated under reduced pressure. Thecrude product was purified by flash chromatography on silica gel(DCM/methanol gradient) and preparative HPLC (SunFire C18 column,CH₃CN/1% TFA in H₂O gradient, pure fractions were treated with DCM and asaturated aqueous NaHCO₃ solution; the combined organic extracts weredried with MgSO₄, filtered and concentrated under reduced pressure) toprovide the title compound as a yellow solid.

HPLC Rt_(M10)=2.27 min; ESIMS: 461 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.46-8.41 (m, 1H), 8.30-8.26 (m, 1H),7.66-7.59 (m, 2H), 6.77-6.72 (m, 1H), 6.37-6.29 (m, 1H), 6.14-6.07 (m,1H), 4.90-4.79 (m, 1H), 4.25-4.11 (m, 3H), 3.99 (s, 3H), 3.98-3.78 (m,1H), 3.70-3.41 (m, 7H), 2.10-1.93 (m, 2H).

alternative method f2: the carboxylic acid, HOBT, EDC and DMF werereplaced with the carboxylic acid chloride and DCM

alternative method f3: the carboxylic acid, HOBT, EDC and DMF werereplaced with a chloroformate and DCM

alternative method f4: the carboxylic acid, HOBT and EDC were replacedwith a carbamic chloride

Examples D2 to D40

The compounds listed in Table 4 were prepared by a procedure analogousto that used in Example D1.

TABLE 4 HPLC Rt MS [min] [m/z; Example Compound (method) (M + 1)⁺] D2

2.07 (M10) 455 D3

2.08 (M10) 413 D4

2.72 (M10) 465 D5

2.61 (M10) 506 D6

2.22 (M10) 438 D7

2.14 (M10) 480 D8

2.14 (M10) 480 D9

3.19 (M10) 451 D10

3.06 (M10) 437 D11

2.96 (M10) 411 D12

2.63 (M10) 425 D13

3.22 (M10) 463 D14

2.23 (M10) 478 D15

2.67 (M10) 439 D16

1.62 (M8) 513 D17

1.55 (M13) 488 D18

1.75 (M13) 448 D19

1.56 (M13) 488 D20

1.60 (M8) 474 D21

1.50 (M13) 474 D22

3.01 (M10) 467 D23

3.11 (M10) 411 D24

1.63 (M9) 467 D25

1.66 (M9) 411 D26

2.31 (M10) 463 D27

2.62 (M10) 461 D28

2.68 (M10) 451 D29

2.68 (M10) 451 D30

2.63 (M10) 440 D31

2.24 (M10) 461 D32

1.69 (M13) 448 D33

1.80 (M13) 462 D34

1.77 (M13) 466 D35

1.90 (M13) 493 D36

2.01 (M13) 493 D37

1.28 (M13) 475 D38

2.19 (M10) 464 D39

2.24 (M10) 450 D40

2.24 (M10) 450

Example E1{(S)-3-[1-(6-Methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone(according to Scheme 5)

a) 7-Chloro-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine

A mixture of 7-chloro-1H-pyrido[3,4-b][1,4]oxazin-2-one (CAS registry928118-43-8) (630 mg, 3.41 mmol) and BH₃*THF (1 M in THF) (10.2 ml, 10.2mmol) in THF (20 ml) was stirred for 2 h at 80° C. The reaction mixturewas quenched with MeOH, NaOH aq. solution 1 M was added and the mixturewas extracted with EtOAc. Combined organic layers were washed withbrine, dried over Na₂SO₄, filtered and evaporated. The crude product waspurified by flash chromatography on silica gel (heptane/EtOAc 100:0 to50:50 in 12 min) to provide the title compound as a white solid (432 mg,74% yield).

HPLC Rt_(M1)=0.47 min; ESIMS: 171 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.74 (s, 1H), 6.46 (s, 1H), 4.43 (br s, 1H)4.21-4.25 (m, 2H), 3.48-3.51 (m, 2H).

b)7-Chloro-1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]-oxazine

A mixture of 7-chloro-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine (127 mg,0.74 mmol), 5-bromo-2-methoxy-3-methylpyridine (CAS registry760207-87-2) (0.196 g, 0.986 mmol), Cs₂CO₃ (534 mg, 1.64 mmol) and XPhos(28 mg, 0.06 mmol) in dioxane (3.5 ml) was degassed with argon andPd₂(dba)₃ (27 mg, 0.03 mmol) was added. After stirring for 3.5 h at 100°C. the reaction mixture was filtered over hyflo, sat. aq. NaHCO₃ soln.was added and the mixture was extracted with EtOAc. Combined organiclayers were washed with brine, dried over Na₂SO₄, filtered andevaporated. The crude product was purified by flash chromatography onsilica gel (heptane/EtOAc 95:5 to 40:60 in 14 min) to provide the titlecompound as a pale colored solid (190 mg, 87% yield).

HPLC Rt_(M1)=1.04 min; ESIMS: 292 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.94 (d, 1H), 7.80 (s, 1H), 7.31 (d, 1H),6.31 (s, 1H), 4.34-4.37 (m, 2H), 4.01 (s, 3H), 3.68-3.72 (m, 2H), 2.24(s, 3H).

c)1-(6-Methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-ol

A mixture of7-chloro-1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine(190 mg, 0.65 mmol), tetramethyl-t-butyl-XPhos (13 mg, 0.03 mmol) indioxane (3 ml) and 5M aq. KOH soln. (0.04 ml, 1.95 mmol) was degassedwith argon and Pd₂(dba)₃ (6 mg, 0.01 mmol) was added. After stirring for17.5 h at 100° C. the reaction mixture was filtered over hyflo, thefiltrate was dried over Na₂SO₄, filtered and evaporated. The crudeproduct was purified by flash chromatography on silica gel (EtOAc/MeOH100:0 to 85:15 in 17 min) to provide the title compound as a white solid(111 mg, 62% yield).

HPLC Rt_(M1)=0.67 min; ESIMS: 274 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 10.32 (brs, 1H), 8.02 (d, 1H), 7.62 (m, 1H),6.89 (s, 1H), 4.84 (s, 1H), 4.17-4.21 (m, 2H), 3.91 (s, 3H), 3.61-3.66(m, 2H), 2.17 (s, 3H).

d)(S)-3-[1-(6-Methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

A solution of1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-ol(111 mg, 0.41 mmol) in DMF (3 ml) was treated with NaH (33 mg, 0.81mmol) for 10 min at 20° C.(R)-3-Methanesulfonyloxy-pyrrolidine-1-carboxylic acid tert-butyl ester(CAS registry 127423-61-4) (162 mg, 0.61 mmol) was added. After stirringfor 19 h at 60° C. and 18 h at 80° C. sat. aq. NaHCO₃ soln. was addedand the reaction mixture was extracted with TBME. Combined organiclayers were washed with brine, dried over Na₂SO₄, filtered andevaporated. The crude product was purified by flash chromatography onsilica gel (heptane/EtOAc 93:7 to 40:60 in 13.5 min) to provide thetitle compound as a pale yellow oil (107 mg, 59% yield).

HPLC Rt_(M1)=1.21 min; ESIMS: 443 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.61 (br s, 1H), 7.32 (br s1H), 5.71 (s, 1H), 5.41 (br s, 1H), 4.32 (br s, 2H), 3.99 (s, 3H),3.65-3.70 (m, 2H), 3.37-3.61 (m, 4H), 2.23 (s, 3H), 1.58 (s, 9H),0.82-0.97 (m, 2H).

e)1-(6-Methoxy-5-methyl-pyridin-3-yl)-7-((S)-pyrrolidin-3-yloxy)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine

A solution of(S)-3-[1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]-oxazin-7-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (103 mg, 0.23 mmol) and TFA (0.179 ml, 2.33 mmol)in DCM (1.8 ml) was stirred for 18 h at rt. Sat. aq. Na₂CO₃ soln. wasadded and the reaction mixture was extracted with DCM. Combined organiclayers were washed with brine, dried over Na₂SO₄, filtered andevaporated, the title compound was as a pale yellow foam (72 mg, 90%yield).

HPLC Rt_(M1)=0.64 min; ESIMS: 343 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.62 (s, 1H), 7.32 (m, 1H),5.70 (s, 1H), 5.29-5.35 (m, 1H), 4.29-4.33 (m, 2H), 3.99 (s, 3H),3.65-3.69 (m, 2H), 2.82-3.14 (m, 4H), 2.22 (s, 3H), 1.80-2.10 (m, 2H).

f){(S)-3-[1-(6-Methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone

A mixture of 1-methyl-1H-imidazole-4-carboxylic acid (CAS registry41716-18-1) (15 mg, 0.12 mmol), HBTU (53 mg, 0.14 mmol) and DIPEA (0.025ml, 0.14 mmol) in DMF (0.6 ml) was stirred at rt for 5 min. A solutionof1-(6-methoxy-5-methyl-pyridin-3-yl)-7-((S)-pyrrolidin-3-yloxy)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine(0.037 g, 0.11 mmol) in DMF (0.6 ml) was added. After stirring for 20 hat rt water was added and the reaction mixture and was extracted withEtOAc. Combined organic layers were washed with brine, dried overNa₂SO₄, filtered and evaporated. The crude product was purified by prep.RP-HPLC (column SunFire C18, H₂O+0.1% TFA/ACN+0.1% TFA 90:10 to 60:40 in16 min) to provide the title compound as a pale yellow foam (24 mg, 49%yield).

HPLC Rt_(M1)=0.74 min; ESIMS: 451 [(M+H)⁺].

¹H NMR (400 MHz, DMSO): δ 8.00 (m, 1H), 7.58-7.63 (m, 3H), 7.53 (d, 1H),5.51 (d, 1H), 5.29-5.40 (m, 1H), 4.23-4.29 (m, 2H), 3.99 (s, 3H),3.77-4.19 (m, 2H), 3.66 (m, 5H), 3.39-3.63 (m, 2H), 2.15 (s, 3H),1.89-2.11 (m, 2H).

Examples E2 to 11

The compounds listed in Table 5 were prepared by a procedure analogousto that used in Example E1.

TABLE 5 HPLC Rt MS Compound/ [min] [m/z; Example Reaction Conditions(method) (M + 1)⁺] E2

1.19 (M1) 443 (S)-3-[1-(6-Methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]- pyrrolidine-1-carboxylicacid tert-butyl ester Buchwald amination condition: CA4 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8, IA9,127423-61-4 E3

0.84 (M1) 455 {(S)-3-[1-(6-Methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone Buchwald aminationcondition: CA4 Amide bond condition: CB6 Side chain introductioncondition: CC1 Precursors used: CAS 928118-43-8, 127423-61-4, IA9, acylchloride 40191-32-0 E4

1.09 (M1) 328 {(S)-3-[1-(6-Difluoromethoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-methanone Buchwald amination condition: CA4Amide bond condition: CB4 Side chain introduction condition: CC1Precursors used: CAS 928118-43-8, IA8, 127423-61-4, 64096-87-3 E5

0.79 (M1) 523 {(S)-3-[1-(5-Chloro-6-methoxy-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-methanone Buchwald amination condition: CA4Amide bond condition: CB1 Side chain introduction condition: CC1Precursors used: CAS 928118-43-8, 127423-61-4, IA11, 64096-87-3 E6

0.92 (M1) 557 (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[1-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald amination condition: CA4Amide bond condition: CB1 Side chain introduction condition: CC1Precursors used: CAS 928118-43-8/127423- 61-4/IA21/64096-87-3 E7

0.87 (M1) 505 {(S)-3-[1-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4- yl)-methanone Buchwaldamination condition: CA4 Amide bond condition: CB1 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8/127423-61-4/IA21/41716-18-1 E8

0.75 (M1) 471 {(S)-3-[1-(5-Chloro-6-methoxy-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone Buchwaldamination condition: CA4 Amide bond condition: CB1 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8,127423-61-4, IA11, 41716-18-1 E9

0.95 (M1) 491 {(S)-3-[1-(6-Difluoromethoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone Buchwaldamination condition: CA4 Amide bond condition: CB1 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8/127423-61-4/IA8/5337-03-1 E10

0.84 (M1) 487 {(S)-3-[1-(6-Difluoromethoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4- yl)-methanone Buchwaldamination condition: CA4 Amide bond condition: CB1 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8/127423-61-4/IA8/41716-18-1 E11

1.01 (M1) 447 Cyclopropyl-{(S)-3-[1-(6-difluoromethoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald aminationcondition: CA4 Amide bond condition: CB6 Side chain introductioncondition: CC1 Precursors used: CAS 928118-43-8/127423- 61-4/IA8/Acylchloride: 4023-34-1

Reference Examples E12 to E13

The compounds listed in Table 5a were prepared by a procedure analogousto that used in Example E1, applying adequate protecting groupstrategies.

TABLE 5a HPLC Rt MS Reference Compound/ [min] [m/z; Example ReactionConditions (method) (M + 1)⁺] E12

0.54 (M16) 489 (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[1-(6-hydroxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald amination condition: CA2Amide bond condition: CB7 Side chain introduction condition: CC1Precursors used: CAS 928118-43-8/127423- 61-4/IA69/64096-87-3 E13

0.63 (M16) 519 (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[1-(5-hydroxymethyl-6-methoxy-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald aminationcondition: CA2 Amide bond condition: CB7 Side chain introductioncondition: CC1 Precursors used: CAS 928118-43-8/127423-61-4/IA70/64096-87-3

Example F1(1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone(according to Scheme 6)

a) 7-Chloro-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine

A solution of 7-chloro-1H-pyrido[3,4-b][1,4]oxazin-2-one (CAS registry928118-43-8) (3.70 g, 20 mmol) in THF (63 ml) was treated with BH₃*THF(1M in THF, 47 ml, 47 mmol). The reaction mixture was stirred at 75° C.for 1 h, then cooled down to rt and quenched with methanol (24 ml, 600mmol). The reaction mixture was concentrated under reduced pressure andthe residue was taken up with EtOAc and washed with sat. aq. NaHCO₃soln. The organic layer was dried over MgSO₄, filtered and concentratedunder reduced pressure to afford the title product as a pale yellowsolid (3.3 g, 96% yield).

UPLC Rt_(M1)=0.47 min; ESIMS: 171 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.53 (s, 1H), 7.11 (br s, 1H), 6.47 (s,1H), 4.09 (t, 2H), 3.17-3.38 (m, 2H).

b) 2,3-Dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-ol

A mixture of 7-chloro-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine (1.08 g,6.33 mmol), aq. KOH soln. (1.07 g, 19 mmol KOH in 5.4 ml water),2-di-t-butylphosphino-3,4,5,6-tetramethyl-2′,4′,6-tri-i-propylbiphenyl98% (0.30 g, 0.63 mmol) and Pd₂(dba)₃ (0.29 g, 0.32 mmol) in dioxane(32.5 ml) was degassed three times with nitrogen, the tube was sealedand the reaction mixture was stirred at 100° C. for 5 h. After coolingto rt, the reaction mixture was filtered through hyflo, rinsed withEtOAc and methanol. The filtrates were concentrated and the titlecompound was obtained after flash chromatography on silica gel(DCM/MeOH, 98:2 to 75:25) as an orange residue (660 mg, 69% yield)

UPLC Rt_(M1)=0.34 min; ESIMS: 153 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 10.33 (br s, 1H), 7.03 (br s, 1H), 6.71 (s,1H), 5.15 (s, 1H), 3.95 (t, 2H), 3.25 (m, 2H).

c)(S)-3-(2,3-Dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

A dry solution of 2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-ol (0.66 g,4.34 mmol) and (R)-3-methanesulfonyloxy-pyrrolidine-1-carboxylic acidtert-butyl ester (CAS registry 127423-61-4) (1.73 g, 6.51 mmol) in DMF(40 ml) was treated with sodium hydride (60% in mineral oil, 0.21 g,8.68 mmol) and the reaction mixture was stirred at 80° C. for 18 h.After cooling to rt, the reaction mixture was diluted with TBME andwashed with sat. aq. NaHCO₃ soln. The organic layer was dried overMgSO₄, filtered, concentrated and the title compound was obtained afterflash chromatography on silica gel (cyclohexane/EtOAc, 95:5 to 30:70) asa yellow oil (1.035 g, 75% purity, 56% yield)

UPLC Rt_(M1)=0.65 min; ESIMS: 322 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.54 (s, 1H), 5.86 (s, 1H), 5.42 (br s, 1H),4.25-4.41 (m, 1H), 4.19 (t, 2H), 3.38-3.66 (m, 6H), 2.00-2.18 (m, 2H),1.46 (d, 9H).

d)(S)-3-[1-(6-Methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

A mixture of(S)-3-(2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (254 mg, 0.79 mmol),5-bromo-2-methoxy-3-methylpyridine (CAS registry 760207-87-2) (208 mg,1.03 mmol), XPhos (30 mg, 0.06 mmol), and NaOtBu (167 mg, 1.74 mmol) indioxane (6 ml) was degassed with argon for 5 min, then Pd₂(dba)₃ (29 mg,0.03 mmol) was added. The tube was filled with argon, sealed and thereaction mixture was stirred at 100° C. for 2 h. After cooling to rt,the reaction mixture was filtered through hyflo, rinsed with EtOAc andthe filtrates were washed with sat. aq. NaHCO₃ soln. The aq. layer wastwice reextracted with EtOAc, the combined organic layers were driedover Na₂SO₄, filtered, concentrated and the title compound was obtainedafter flash chromatography on silica gel (heptane/EtOAc, 100:0 to 50:50)as a clear gum (274 mg, 78% yield). UPLC Rt_(M1)=1.20 min; ESIMS: 443[(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.61 (br s, 1H), 7.30-7.35 (m,1H), 5.71 (s, 1H), 5.34-5.46 (m, 1H), 4.31 (br s, 2H), 3.99 (s, 3H),3.68 (t, 2H), 3.34-3.62 (m, 4H), 2.23 (s, 3H), 2.01-2.09 (m, 2H), 1.44(s, 9H).

e)1-(6-Methoxy-5-methyl-pyridin-3-yl)-7-((S)-pyrrolidin-3-yloxy)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine

A solution of(S)-3-[1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (364 mg, 0.82 mmol) in DCM (6 ml) was treated withTFA (0.63 ml, 8.23 mmol) and the reaction mixture was stirred at rt for18 h, then quenched with sat. aq. NaHCO₃ soln. and extracted with DCM.The organic layer was dried over MgSO₄, filtered and concentrated underreduced pressure to afford the title product as a red oil, which wasused in the next step without further purification (313 mg, 90% purity,quantitative yield).

UPLC Rt_(M1)=0.65 min; ESIMS: 343 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.93 (d, 1H), 7.62 (s, 1H), 7.32 (d, 1H),5.70 (s, 1H), 5.26-5.36 (m, 1H), 4.31 (t, 2H), 3.99 (s, 3H), 3.67 (t,2H), 2.95-3.15 (m, 3H), 2.81-2.92 (m, 1H), 2.22 (s, 3H), 1.98-2.10 (m,1H), 1.79-1.90 (m, 1H).

f)(1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl)}-methanone

A solution of 1,1-dioxo-hexahydro-1lambda*6*-thiopyran-4-carboxylic acid(CAS registry 64096-87-3) (106 mg, 0.59 mmol) in DMF (4 ml) was treatedwith HBTU (225 mg, 0.59 mmol) and DIPEA (0.24 ml, 1.37 mmol). Theresulting orange solution was stirred at rt for 5 min, then a solutionof1-(6-methoxy-5-methyl-pyridin-3-yl)-7-((S)-pyrrolidin-3-yloxy)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine(156 mg, 0.46 mmol) in DMF (2 ml) was added. The reaction mixture wasstirred at rt for 1 h then concentrated under reduced pressure and theresidue was taken up with DCM and washed with sat. aq. NaHCO₃ soln. Theorganic layer was dried by passing it through a phase separatingcartridge, concentrated and the title compound was obtained after SFCchromatography (column DEAP (250 mm×30 mm, 60 A, 5 μm) Princeton,gradient 11-16% of methanol in supercritical CO₂ in 6 min) as a slightlycoloured solid (112 mg, 49% yield).

UPLC Rt_(M1)=0.81 min; ESIMS: 503 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.01 (s, 1H), 7.61 (m, 1H), 7.52 (d, 1H),5.52 (d, 1H), 5.24-5.43 (m, 1H), 4.26 (br s, 2H), 3.89 (s, 3H),3.59-3.79 (m, 3H), 3.41-3.56 (m, 2H), 3.21-3.39 (m, 1H), 2.98-3.21 (m,4H), 2.67-2.83 (m, 1H), 1.84-2.20 (m, 9H).

¹H NMR (600 MHz, DMSO-d₆): δ 8.01 (s, 1H), 7.63-7.59 (m, 1H), 7.55-7.51(m, 1H), 5.55-5.51 (m, 1H), 5.43-5.24 (m, 1H), 4.29-4.22 (m, 2H), 3.90(s, 3H), 3.80-3.60 (m, 2H), 3.56-3.37 (m, 3H), 3.28-2.99 (m, 5H),2.89-2.66 (m, 1H), 2.19-2.09 (m, 4H), 2.08-1.98 (m, 2H), 1.98-1.86 (m,3H).

Crystallization of Example F1 by Heating and Cooling inIsopropanol/Diethyl Ether

474 mg of amorphous Example F1 was suspended in 1.4 mL of isopropanol.The mixture was heated to 70° C. and stirred at 70° C. to allow completedissolution of Example F1. The solution was cooled down to RT, a glueresidue was formed. 2 mL of diethyl ether was added and the slurry wasstirred for 48 h. A white suspension was formed. The suspension wasfiltered and the solid was dried at 40° C., 15 mbar. A fine, whitepowder was obtained. The material contains only slight residual solvent(<0.5%). A crystalline anhydrous form of Example F1 with an onsetmelting of 148.77° C. was obtained.

List of most significant 2-Theta peaks from X-ray Powder DiffractionPattern with tolerances±0.5 of Example F1 anhydrous form (Method M1)(including low/weak peaks for information). Note: This list of peaks isnot exhaustive but are only “inter alia”.

2-Theta in deg Intensity 9.1 Low 10.2 Medium 11.9 Medium 13.0 Low 17.1Strong 17.7 Medium, unresolved 18.7 Medium 20.3 Medium, unresolved 20.8Medium, unresolved 26.0 Medium/low 26.7 Medium 23.2 Medium/low 24.1Medium/low 24.8 Medium/low 29.3 Medium/low 27.4 Medium/low 21.4Medium/low

Examples F2 to F15

The compounds listed in Table 6 were prepared by a procedure analogousto that used in Example F1.

TABLE 6 HPLC Rt MS Compound/ [min] [m/z; Example Reaction Conditions(method) (M + 1)⁺] F2

0.79 (M1) 507 (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[1-(5-fluoro-6-methoxy-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald amination condition: CA2 Amide bondcondition: CB2 Side chain introduction condition: CC1 Precursors used:CAS 928118-43-8/127423- 61-4/IA10/64096-87-3 F3

0.73 (M1) 455 {(S)-3-[1-(5-Fluoro-6-methoxy-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)- methanone Buchwaldamination condition: CA2 Amide bond condition: CB2 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8/127423-61-4/IA10/41716-18-1 F4

0.83 (M1) 466 2-Methoxy-5-{7-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-pyrido[3,4-b][1,4]oxazin-1-yl}-nicotinonitrile Buchwald aminationcondition: CA2 Amide bond condition: CB6 Side chain introductioncondition: CC1 Precursors used: CAS 928118-43-8, 127423-61-4, IA12, acylchloride 40191-32-0 F5

0.73 (M1) 462 2-Methoxy-5-{7-[(S)-1-(1-methyl-1H-imidazole-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-pyrido[3,4-b][1,4]oxazin-1-yl}-nicotinonitrile Buchwald aminationcondition: CA2 Amide bond condition: CB1 Side chain introductioncondition: CC1 Precursors used: CAS 928118-43-8, 127423-61-4, IA12,41716-18-1 F6

0.76 (M1) 503 {(S)-3-[1-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)- methanone Buchwaldamination condition: CA4 Amide bond condition: CB6 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8,127423-61-4, IA1, acyl chloride 40191-32-0 F7

0.82 (M1) 539 {(S)-3-[1-(5-Difluoromethyl-6-methoxy-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-methanone Buchwald amination condition: CA2Amide bond condition: CB2 Side chain introduction condition: CC1Precursors used: CAS 928118-43-8/127423- 61-4/IA6/64096-87-3 F8

0.88 (M1) 491 {(S)-3-[1-(5-Difluoromethyl-6-methoxy-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)- methanone Buchwaldamination condition: CA2 Amide bond condition: CB6 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8/127423-61-4/IA6/Acyl chloride 40191-32-0 F9

0.83 (M1) 451 1-{(S)-3-[1-(5-Difluoromethyl-6-methoxy-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-2-methoxy-ethanone Buchwald amination condition:CA2 Amide bond condition: CB6 Side chain introduction condition: CC1Precursors used: CAS 928118-43-8/127423- 61-4/IA6/Acyl chloride:38870-89-2 F10

0.77 (M1) 487 {(S)-3-[1-(5-Difluoromethyl-6-methoxy-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4- yl)-methanone Buchwaldamination condition: CA2 Amide bond condition: CB3 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8/127423-61-4/IA6/41716-18-1 F11

0.76 (M1) 499 1-{(S)-3-[1-(5-Difluoromethyl-6-methanesulfonyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}- 2-methoxy-ethanone Buchwaldamination condition: CA1 Amide bond condition: CB6 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8/127423-61-4/IA4/Acyl chloride: 38870-89-2 F12

0.81 (M1) 503 (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{3-[1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]- pyrrolidin-1-yl}-methanoneBuchwald amination condition: CA4 Amide bond condition: CB1 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8/141699-57-2/IA9/64096-87-3 F13

1.12 (M1) 454 5-{7-[(S)-1-(1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-pyrido[3,4-b][1,4]oxazin-1-yl}-2- methoxy-nicotinonitrileBuchwald amination condition: CA2 Amide bond condition: CB1 Side chainintroduction condition: CC1 Precursors used: CAS 928118-43-8,127423-61-4, IA12, 64096-87-3 F14

0.89 (M1) 35.9 (CD12) 521 (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(R)-3-fluoro-4-[1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald aminationcondition: CA2 Amide bond condition: CB4 Side chain introductioncondition: CC1 Precursors used: CAS 869481-93-6/1174020-51-9 Chiralseparation method: CD12 F15

0.89 (M1) 45.8 (CD12) 521 (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(R)-3-fluoro-4-[1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald aminationcondition: CA2 Amide bond condition: CB4 Side chain introductioncondition: CC1 Precursors used: CAS 869481-93-6/1174020-51-9 Chiralseparation method: CD12

Example G1Imidazo[2,1-b]thiazol-6-yl-{(S)-3-[1-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone

a) 7-Bromo-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine

A solution of 7-bromo-1H-pyrido[3,4-b][1,4]oxazin-2-one (CAS registry943995-72-0) (2.93 g, 12.79 mmol) in THF (40 ml) was treated withBH₃*THF (1M in THF, 30 ml, 30 mmol). The reaction mixture was stirred at80° C. for 1.5 h, then cooled down to rt and quenched with methanol. Thereaction mixture was concentrated under reduced pressure and the residuewas taken up with EtOAc and washed with aq. 1M NaOH soln. The organiclayer was dried over Na₂SO₄, filtered and concentrated under reducedpressure to afford the title product as a white solid. (2.48 g, 90%yield).

UPLC Rt_(M1)=0.49 min; ESIMS: 217 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.72 (s, 1H), 6.60 (s, 1H), 4.42 (br s, 1H),4.20-4.24 (m, 2H), 3.49 (m, 2H).

b) 7-Bromo-2,3-dihydro-pyrido[3,4-b][1,4]oxazine-1-carboxylic acidbenzyl ester

A dry solution of 7-bromo-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine (1.85g, 8.60 mmol) in THF (50 ml) was portionwise treated at 0° C. with 60%NaH in mineral oil (0.52 g, 12.90 mmol) and the reaction mixture wasstirred at 0° C. for 1 h. Benzyl chloroformate (CAS registry 501-53-1)(1.40 ml, 9.85 mmol) was dropwise added and the reaction mixture wasallowed to warm to rt and to stir for 20 h, finally quenched withmethanol and then diluted with sat. aq. NaHCO₃ soln. and extracted withEtOAc. The organic layer was dried over Na₂SO₄, filtered, concentratedand the title compound was obtained after flash chromatography on silicagel (heptane/EtOAc, 100:0 to 60:40) as a white solid (2.06 g, 68%yield).

UPLC Rt_(M1)=1.14 min; ESIMS: 349 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.28 (br s, 1H), 7.98 (s, 1H), 7.35-7.46 (m,5H), 5.30 (s, 2H), 4.20-4.27 (m, 2H), 3.92-4.01 (m, 2H).

c) 2,3-Dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-ol

A mixture of 7-bromo-2,3-dihydro-pyrido[3,4-b][1,4]oxazine-1-carboxylicacid benzyl ester (1.27 g, 3.63 mmol), aq. KOH soln. (0.90 g, 16 mmolKOH in 3.2 ml water),2-di-t-butylphosphino-3,4,5,6-tetramethyl-2′,4′,6-tri-i-propylbiphenyl98% (0.26 g, 0.54 mmol) in dioxane (16 ml) was degassed with argon for 5min, then Pd₂(dba)₃ (0.25 g, 0.27 mmol) was added. The tube was filledwith argon, then sealed and the reaction mixture was stirred at 100° C.for 19 h. After cooling to rt, the reaction mixture was filtered throughhyflo, rinsed with EtOAc and methanol. The filtrates were dried overNa₂SO₄, filtered, concentrated and the title compound was obtained afterflash chromatography on silica gel (DCM/MeOH, 95:5 to 60:40) as anorange residue (262 mg, 47% yield).

UPLC Rt_(M1)=0.32 min; ESIMS: 153 [(M+H)⁺]

¹H NMR (400 MHz, DMSO-d₆): δ 10.33 (br.s, 1H), 7.03 (br.s, 1H), 6.71 (s,1H), 5.15 (s, 1H), 3.95 (t, 2H), 3.25 (td, 2H).

d)(S)-3-(2,3-Dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

A dry solution of 2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-ol (200 mg,0.66 mmol) and (R)-3-methanesulfonyloxy-pyrrolidine-1-carboxylic acidtert-butyl ester (CAS registry 127423-61-4) (262 mg, 0.99 mmol) in DMF(6 ml) was treated with sodium hydride 60% in mineral oil (53 mg, 1.33mmol) and the reaction mixture was stirred at 80° C. for 17 h. Aftercooling to rt, the reaction mixture was diluted with TBME and washedwith sat. aq. NaHCO₃ soln. The organic layer was dried over Na₂SO₄,filtered, concentrated and the title compound was obtained after flashchromatography on silica gel (heptane/EtOAc, 88:12 to 0:100) as an oil(140 mg, 66% yield).

UPLC Rt_(M1)=0.66 min; ESIMS: 322 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.54 (s, 1H), 5.86 (s, 1H), 5.42 (br.s, 1H),4.25-4.41 (m, 1H), 4.19 (t, 2H), 3.38-3.66 (m, 6H), 2.00-2.18 (m, 2H),1.46 (d, 9H).

e)(S)-3-[1-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

A mixture of(S)-3-(2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (115 mg, 0.36 mmol),5-bromo-2-methoxy-3-trifluoromethylpyridine (CAS registry 1214377-42-0)(119 mg, 0.47 mmol), XPhos (14 mg, 0.03 mmol), and NaOtBu (76 mg, 0.79mmol) in dioxane (2.5 ml) was degassed with argon for 5 min, thenPd₂(dba)₃ (13 mg, 0.01 mmol) was added. The tube was filled with argon,then sealed and the reaction mixture was stirred at 100° C. for 2 h.After cooling to rt, the reaction mixture was filtered through hyflo,rinsed with EtOAc and the filtrates were washed with sat. aq. NaHCO₃soln. The organic layer was dried over Na₂SO₄, filtered, concentratedand the title compound was obtained after flash chromatography on silicagel (heptane/EtOAc, 93:7 to 40:60) as a clear gum. (91 mg, 51% yield).

UPLC Rt_(M1)=1.27 min; ESIMS: 497 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.29 (d, 1H), 7.79 (d, 1H), 7.64 (d, 1H),5.70 (s, 1H), 5.36-5.46 (m, 1H), 4.34 (br s, 2H), 4.09 (s, 3H), 3.70 (t,2H), 3.34-3.62 (m, 4H), 2.02-2.11 (m, 2H), 1.44 (s, 9H).

f)1-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-7-((S)-pyrrolidin-3-yloxy)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine

A solution of(S)-3-[1-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (88 mg, 0.18 mmol) in DCM (1.3 ml) was treatedwith TFA (0.14 ml, 1.77 mmol) and the reaction mixture was stirred at rtfor 17 h, then quenched with sat. aq. Na₂CO₃ soln. and extracted withDCM. The organic layer was dried over Na₂SO₄, filtered and concentratedunder reduced pressure to afford the title compound (66 mg, 94% yield).

UPLC Rt_(M1)=0.72 min; ESIMS: 397 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.28 (d, 1H), 7.79 (d, 1H), 7.65 (s, 1H),5.68 (s, 1H), 5.31-5.39 (m, 1H), 4.31-4.37 (m, 2H), 4.08 (s, 3H),3.67-3.72 (m, 2H), 3.01-3.18 (m, 3H), 2.85-2.97 (m, 1H), 2.01-2.13 (m,1H), 1.82-1.95 (m, 1H).

g)Imidazo[2,1-b]thiazol-6-yl-{(S)-3-[1-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone

A solution of imidazo[2,1-b]thiazole-6-carboxylic acid, hydrobromide(1:1) (CAS registry 725234-39-9) (25 mg, 0.10 mmol) in DMF (0.45 ml) wastreated with HBTU (41 mg, 0.11 mmol) and DIPEA (0.04 ml, 0.21 mmol). Theresulting orange solution was stirred at rt for 5 min, then a solutionof1-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-7-((S)-pyrrolidin-3-yloxy)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine(32 mg, 0.08 mmol) in DMF (0.45 ml) was added. The reaction mixture wasstirred at rt for 17 h, then concentrated under reduced pressure and theresidue was taken up with EtOAc and washed with brine. The organic layerwas dried over Na₂SO₄, filtered, concentrated and the title compound wasobtained after RP prep.

HPLC (Sunfire PrepC18 30×100 mm, 5 μm; solvent A: H₂O+0.1 Vol.-% TFA;solvent B: CH₃CN+0.1 Vol.-% TFA, gradient 15-45% B in 16 min). Afterfiltration over an Agilent PL-HCO₃ MP SPE cartridge, the title compoundwas obtained as a solid (23 mg, 52% yield).

UPLC Rt_(M1)=1.00 min; ESIMS: 547 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.49 (dd, 1H), 8.16-8.20 (m, 2H), 7.92 (dd,1H), 7.57 (d, 1H), 7.37 (dd, 1H), 5.63 (d, 1H), 5.33-5.45 (m, 1H),4.25-4.31 (m, 2H), 3.52-4.14 (m, 9H), 1.88-2.12 (m, 2H).

Examples G2 to G3

The compounds listed in Table 7 were prepared by a procedure analogousto that used in Example G1.

TABLE 7 HPLC Rt MS Compound/ [min] [m/z; Example Reaction Conditions(method) (M + 1)⁺] G2

0.80 (M1)  503 (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(R)-3-[1-(6-methoxy-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald amination condition: CA2 Amide bondcondition: CB1 Side chain introduction condition: CC1 Precursors used:CAS 943995-72-0/132945- 75-6/IA9/64096-87-3 G3

0.86 (M1M1) 520 (5-Amino-1-methyl-1H-imidazol-4-yl)-{(S)-3-[1-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]- pyrrolidin-1-yl}-methanoneBuchwald amination condition: CA2 Amide bond condition: CB1 Side chainintroduction condition: CC1 Precursors used: CAS 943995-72-0/127423-61-4/IA21)/IB3)/Product obtained after Deboc reaction using TFA inCH₂Cl₂ done in conventional way

Examples H1 to H16

The compounds listed in Table 8 were prepared by chromatographicdiastereomer separation.

TABLE 8 HPLC Rt MS Compound/ [min] [m/z; Example Reaction Conditions(method) (M + 1)⁺] H1

  (1,1-Dioxo-tetrahydro-1lambda*6*-thiophen-3-yl)-{(S)-3-[1-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald aminationcondition: CA6 Amide bond condition: CB4 Side chain introductioncondition: CC2 Precursors used: IA1, CAS 127423-61-4/64096-87-3 Chiralseparation method: CD5 0.87 (M2) 531 H2

  (1,1-Dioxo-tetrahydro-1lamdba*6*-thiophen-3-yl)-{(S)-3-[1-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-7-yloxy]-pyrrolidin-1-yl}-methanone Buchwald aminationcondition: CA6 Amide bond condition: CB4 Side chain introductionconditions: CC2 Precursors used: IA1, CAS 127423-61-4/64096-87-3 Chiralseparation method: CD5 0.87 (M2) 536 H3

  [1,4]Dioxan-2-yl-{(S)-3-[4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}- methanone Buchwald aminationcondition: CA6 Amide bond condition: CB4 Side chain introductioncondition: CC2 Precursors used: IA1, CAS 127423-61-4/89364-41-0 Chiralseparation method: CD6 0.88 (M2) 504 H4

  [1,4]Dioxan-2-yl-{(S)-3-[4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}- methanone Buchwald aminationcondition: CA6 Amide bond condition: CB4 Side chain introductioncondition: CC2 Precursors used: IA1), CAS 127423-61-4/89364-41-0 Chiralseparation method: CD6 0.88 (M2) 504 H5

  {(S)-3-[4-(5,6-Dimethoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-[1,4]dioxan-2-yl-methanone Buchwald amination condition: CA6 Amidebond condition: CB4 Side chain introduction condition: CC2 Precursorsused: IA31, CAS 52605-98-8, 127423-61-4/89364-41-0 Chiral separationmethod: CD1 0.91 (M2) 472 H6

  {(S)-3-[4-(5,6-Dimethoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-[1,4]dioxan-2-yl-methanone Buchwald amination condition: CA6 Amidebond condition: CB4 Side chain introduction condition: CC2 Precursorsused: IA31, CAS 52605-98-8, 127423-61-4/89364-41-0 Chiral separationmethod: CD1 0.91 (M2) 472 H7

  {(S)-3-[4-(5,6-Dimethoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(tetrahydro-furan-2-yl)-methanone Buchwald amination condition:CA9 Amide bond condition: CB4 Side chain introduction condition: CC2Precursors used: IA29, CAS 52605-98-8, 127423-61-4/1264293-76-6 Chiralseparation method: CD1 0.91 (M2) 472 H8

  {(S)-3-[4-(5,6-Dimethoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(tetrahydro-furan-2-yl)-methanone Buchwald amination condition:CA6 Amide bond condition: CB4 Side chain introduction condition: CC2Precursors used: IA29, CAS 52605-98-8, 127423-61-4/1264293-76-6 Chiralseparation method: CD1 0.91 (M2) 472 H9

  {(S)-3-[4-(5,6-Dimethoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-tetrahydro-1lambda*6*-thiophen- 3-yl)-methanoneBuchwald amination condition: CA6 Amide bond condition: CB4 Side chainintroduction condition: CC2 Precursors used: IA29, CAS 52605-98-8,127423-61-4/64096-87-3 Chiral separation method: CD1 0.91 (M2) 504 H10

  {(S)-3-[4-(5,6-Dimethoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-tetrahydro-1lambda*6*-thiophen- 3-yl)-methanoneBuchwald amination condition: CA6 Amide bond condition: CB4 Side chainintroduction condition: CC2 Precursors used: IA29, CAS 52605-98-8,127423-61-4/64096-87-3 Chiral separation method: CD1 0.91 (M2) 504 H11

  (1,1-Dioxo-tetrahydro-1lambda*6*-thiophen-3-yl)-{(S)-3-[4-(5-fluoro-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]- pyrrolidin-1-yl}-methanoneBuchwald amination condition: CA6 Amide bond condition: CB5 Side chainintroduction condition: CC2 Precursors used: IA10, CAS 124432-70-8,127423-61-4/64096-87-3 Chiral separation method: CD1 3.26 (M2) 492 H12

  (1,1-Dioxo-tetrahydro-1lambda*6*-thiophen-3-yl)-{(S)-3-[4-(5-fluoro-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]- pyrrolidin-1-yl}-methanoneBuchwald amination condition: CA6 Amide bond condition: CB5 Side chainintroduction condition: CC2 Precursors used: IA10, CAS 124432-70-8,127423-61-4/64096-87-3 Chiral separation method: CD1 3.25 (M2) 492 H13

  {(S)-3-[4-(5-Chloro-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-tetrahydro-1lambda*6*-thiophen- 3-yl)-methanoneBuchwald amination condition: CA6 Amide bond condition: CB5 Side chainintroduction condition: CC2 Precursors used: IA11, CAS 848366-28-9,127423-61-4/64096-87-3 Chiral separation method: CD2 3.52 (M2) 508 H14

  {(S)-3-[4-(5-Chloro-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-tetrahydro-1lambda*6*-thiophen- 3-yl)-methanoneBuchwald amination condition: CA6 Amide bond condition: CB5 Side chainintroduction condition: CC2 Precursors used: IA11, CAS 848366-28-9,127423-61-4/64096-87-3 Chiral separation method: CD2 3.52 (M2) 508 H15

  [1,4]Dioxan-2-yl-{(S)-3-[4-(5-fluoro-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-methanone Buchwald amination condition: CA6Amide bond condition: CB5 Side chain introduction condition: CC2Precursors used: IA10, CAS 124432-70-8, 127423-61-4/89364-41-0 Chiralseparation method: CD1 3.28 (M2) 460 H16

  [1,4]Dioxan-2-yl-{(S)-3-[4-(5-fluoro-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-methanone Buchwald amination condition: CA6Amide bond condition: CB5 Side chain introduction condition: CC2Precursors used: IA10, CAS 124432-70-8, 127423-61-4/89364-41-0 Chiralseparation method: CD1 3.32 (M2) 460

Example I1(1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[5-fluoro-4-(6-methoxy-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-methanone

a) 5-Fluoro-6-methoxy-4H-benzo[1,4]oxazin-3-one

A solution of 6-bromo-5-fluoro-4H-benzo[1,4]oxazin-3-one (CAS registry1029421-36-0) (5.0 g, 20 mmol) in MeOH (10 ml) was treated with sodiummethoxide solution (30% in MeOH, 11.3 ml, 61 mmol) and CuI (0.4 g, 2mmol). After stirring for 20 h at 80° C., the reaction was quenched withsat. aq. NaHCO₃ soln and extracted with EtOAc. The organic layer wasdried over Na₂SO₄, filtered and concentrated under reduced pressure toobtain a pale yellow solid. (2.2 g, 92% yield).

UPLC Rt_(M1)=0.64 min.

¹H NMR (400 MHz, DMSO-d₆): δ 6.74 (m, 2H), 4.53 (s, 2H), 3.79 (s, 3H).

b) 5-Fluoro-6-hydroxy-4H-benzo[1,4]oxazin-3-one

A solution of 5-fluoro-6-methoxy-4H-benzo[1,4]oxazin-3-one (2.0 g, 10mmol) in DCM (50 ml) was treated at 0° C. with boron tribromide (9.6 ml,101 mmol). The reaction mixture was stirred at rt for 17 h, then cooleddown to 0° C. and quenched with methanol. The mixture was concentratedunder reduced pressure and the residue was taken up with EtOAc andwashed with sat. aq. NaHCO₃ soln. The organic layer was washed with 10%aq. Na₂S₂O₄ soln, dried over Na₂SO₄, filtered and concentrated underreduced pressure. The title compound was obtained after flashchromatography on silica gel (hexane/EtOAc, 100:0 to 60:40) as a brownsolid (780 mg, 42% yield).

UPLC Rt_(M1)=0.49 min; ESIMS: 228 [(M+HCOO)⁻].

¹H NMR (400 MHz, DMSO-d₆): δ 11.00 (s, 1H), 6.65 (d, 1H), 6.45 (t, 1H),4.50 (s, 2H).

c) 5-Fluoro-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol

A solution of 5-fluoro-6-hydroxy-4H-benzo[1,4]oxazin-3-one (780 mg, 4.2mmol) in THF (10 ml) was treated with BH₃*THF (1M in THF, 12.8 ml, 12.8mmol). The reaction mixture was stirred at rt for 17 h, then cooled downto 0° C. and quenched with methanol (30 ml). The reaction mixture wasconcentrated under reduced pressure to obtain a brown oil (720 mg,quantitative yield).

UPLC Rt_(M1)=0.54 min; ESIMS: 170 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.95 (s, 1H), 6.45 (d, 1H), 6.00 (t, 1H),4.09 (m, 2H), 3.45 (m, 2H).

d)(S)-3-(5-Fluoro-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

A solution of triphenylphosphine (1.5 g, 5.7 mmol) in THF (20 ml) wastreated at 0° C. with DEAD (0.900 ml, 5.69). The orange solution wasstirred over 10 min at rt, then5-fluoro-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol (740 mg, 4.37 mmol) and(R)-tert-butyl 3-hydroxypyrrolidine-1-carboxylate (1065 mg, 5.69 mmol)were added. The reaction mixture was stirred for 19 h at 60° C. and thenconcentrated under reduced pressure. The title compound was obtainedafter flash chromatography on silica gel (Hexane/EtOAc, 100:0 to 70:30)as a colourless oil (1.1 g, 74% yield).

UPLC Rt_(M1)=1.07 min; ESIMS: 339 [(M+H)⁺]

¹H NMR (400 MHz, DMSO-d₆): δ 6.45 (d, 1H), 6.00 (t, 1H), 5.42 (br.s,1H), 4.25-4.41 (m, 1H), 4.19 (t, 2H), 3.38-3.66 (m, 6H), 2.00-2.18 (m,2H), 1.46 (d, 9H)

e)(S)-3-[5-fluoro-4-(6-methoxy-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

A mixture of(S)-3-(5-fluoro-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (100 mg, 0.296 mmol),5-bromo-2-methoxy-3-methylpyridine (CAS registry 760207-87-2, 179 mg,0.887 mmol), RuPhos (6.90 mg, 0.015 mmol), NaOtBu (85 mg, 0.887 mmol)and (2-dicyclohylphosphino-2′6′-diisopropyl-11′-biphenyl)(2-(2-aminoethyl)phenyl)palladium(II) (12.07 mg, 0.015 mmol)in dioxane (2 ml) were degassed with argon then sealed and the reactionmixture was stirred at 100° C. for 23 h. After cooling to r.t., thereaction mixture was filtered through hyflo, rinsed with EtOAc and thefiltrates were washed with sat. aq. NaHCO₃ soln. The organic layer wasdried over Na₂SO₄, filtered, concentrated and the title compound wasobtained after flash chromatography on silica gel (Hexane/EtOAc, 100:0to 70:30) as a yellow oil (123 mg, 63% yield).

UPLC Rt_(M1)=1.29 min; ESIMS: 460 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.75 (d, 1H), 7.45 (d, 1H), 6.55 (t, 1H),6.35 (d, 1H), 4.75 (m, 1H), 4.15 (t, 2H), 3.84 (s, 3H), 3.60 (t, 2H),3.38-3.66 (m, 4H), 2.12 (s, 3H), 2.00-2.18 (m, 2H), 1.46 (d, 9H).

f)5-Fluoro-4-(6-methoxy-5-methyl-pyridin-3-yl)-6-((S)-pyrrolidin-3-yloxy)-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of(S)-3-[5-fluoro-4-(6-methoxy-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (123 mg, 0.185 mmol) in DCM (2 ml) was treatedwith 4N HCl/dioxane (0.046 ml, 0.185 mmol). The reaction mixture wasstirred at rt for 3 d, then concentrated under reduced pressure toobtain a black oil (100 mg, 79% yield).

UPLC Rt_(M1)=0.73 min; ESIMS: 360 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.75 (d, 1H), 7.45 (d, 1H), 6.80 (m, 1H),6.75 (m, 1H), 5.20 (m, 1H), 4.15 (t, 2H), 3.84 (s, 3H), 3.60 (t, 2H),3.38-3.66 (m, 4H), 2.12 (s, 3H), 2.00-2.18 (m, 2H).

g)(1,1-Dioxo-hexahydro-lambda*6*-thiopyran-4-yl)-{(S)-3-[5-fluoro-4-(6-methoxy-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-methanone

A solution of 1,1-dioxo-hexahydro-1lambda*6*-thiopyran-4-carboxylic acid(CAS registry 64096-87-3) (33.9 mg, 0.15 mmol) in DCM (2 ml) was treatedat rt with Et₃N (0.061 ml, 0.440 mmol) and HATU (55.7 mg, 0.147 mmol).The resulting orange solution was stirred at rt for 20 min, then asolution of5-fluoro-4-(6-methoxy-5-methyl-pyridin-3-yl)-6-((S)-pyrrolidin-3-yloxy)-3,4-dihydro-2H-benzo[1,4]oxazine(100 mg, 0.147 mmol) in DCM (2 ml) was added. The reaction mixture wasstirred at rt for 1.5 h, then diluted with EtOAc and washed with sat.aq. NaHCO₃ soln. The organic layer was dried over Na₂SO₄, filtered,concentrated and the title compound was obtained after prep. RP-HPLC(SunFire C18 column OBD 5 mm 30×100 mm, gradient 25% to 45% ACN in 16min). The fractions were lyophilized and filtered over a PL-HCO₃ MP SPEcartridge to give a brown solid (54 mg, 71% yield).

UPLC Rt_(M1)=0.94 min; ESIMS: 520 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.75 (d, 1H), 7.45 (d, 1H), 6.55 (t, 1H),6.35 (d, 1H), 4.75 (m, 1H), 4.15 (t, 2H), 3.84 (s, 3H), 3.59-3.79 (m,3H), 3.41-3.56 (m, 2H), 3.21-3.39 (m, 1H), 2.98-3.21 (m, 4H), 2.67-2.83(m, 1H), 1.84-2.20 (m, 9H).

Examples I2 to I3

The compounds listed in Table 9 were prepared by a procedure analogousto that used in Example I1.

TABLE 9 HPLC Rt MS Compound/ [min] [m/z; Example Reaction Conditions(method) (M + 1)⁺] I2

  {(S)-3-[4-(5-Difluoromethyl-6-methoxy-pyridin-3-yl)-5-fluoro-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-hexahydro-1lamda*6*-thiopyran-4-yl)-methanone Buchwald amination condition: CA11Amide bond condition: CB3 Side chain introduction condition: CC4Precursor used: IA6, CAS 1254123-51-7/ 127423-61-4,/64096-87-3 0.97 (M2)556 I3

  {(S)-3-[5-Fluoro-4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)- methanone Buchwaldamination condition: CA11 Amide bond condition: CB3 Side chainintroduction condition: Precursors used: IA1, CAS 127423-61-4/Acylchloride 40191-32-0 0.85 (M2) 520

Example J5-{6-[(S)-1-((S)-1-Acetyl-pyrrolidine-3-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]-oxazin-4-yl}-2-methoxy-nicotinonitrile

A solution of2-methoxy-5-{6-[(S)-1-((S)-pyrrolidine-3-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile(Example D39; 23 mg, 0.051 mmol) in DCM (1 ml) was treated with Et₃N(0.014 ml, 10.4 mg, 0.102 mmol). The solution was stirred at rt for 10min, then acetyl chloride (0.0044 ml, 4.87 mg, 0.061 mmol) was added.The reaction mixture was stirred at rt for 1.5 h. Another 2 eq. of Et₃N(0.014 ml, 10.4 mg, 0.102 mmol). and 1 eq. of acetyl chloride ((0.0037ml, 4.06 mg, 0.051 mmol) were added, stirring was continued at rt for1.5 h. The reaction mixture was diluted with DCM and sat. aq. NaHCO₃soln., then passed through a phase separator, the aq. layer was twiceextracted with DCM, the combined org. layers were concentrated to givethe title compound as a yellow oil which was purified by prep. RP-HPLC(column SunFire C18, 10-85% ACN in 20 min). The fractions were extractedwith DCM/NaHCO₃, dried over MgSO₄, concentrated and lyophilized to givethe title compound as a yellow foam (14 mg, 53% yield).

HPLC Rt_(M10)=2.55 min; ESIMS: 492 [(M+H)⁺].

Example K5-{6-[(S)-1-((R)-1-Acetyl-pyrrolidine-3-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-2-methoxy-nicotinonitrile

This example was prepared in analogy to Example J, starting from2-methoxy-5-{6-[(S)-1-((R)-pyrrolidine-3-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile(Example D40).

HPLC Rt_(M10)=2.55 min; ESIMS: 492 [(M+H)⁺].

Example L2-Methoxy-5-{6-[(S)-1-((R)-1-methyl-pyrrolidine-3-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile

A solution of2-methoxy-5-{6-[(S)-1-((R)-pyrrolidine-3-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile(Example D40, 26 mg, 0.058 mmol) in MeOH (1 ml) was treated with a 37%aq. formaldehyde soln. (0.043 ml, 46.9 mg, 0.578 mmol) and acetic acid(0.004 ml, 4.17 mg, 0.0069 mmol). The solution was stirred under argonat rt for 45 min, then NaBH₃CN (5.65 mg of a 90% solid, 0.081 mmol) wasadded. The resulting mixture was stirred at rt for 45 min, diluted withDCM and sat. aq. NaHCO₃ soln. The aq. layer was twice reextracted withDCM, the combined org. layers were dried over MgSO₄ and concentrated togive the crude title compound that was purified by prep. RP-HPLC (columnSunFire C18, gradient 5-75% ACN in 20 min). The fractions were extractedwith DCM/sat. aq. NaHCO₃ soln, dried over MgSO₄, concentrated andlyophilized to give the title compound as a yellow foam (20 mg, 72%yield).

HPLC Rt_(M11)=2.24 min; ESIMS: 464 [(M+H)⁺].

Example M4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-6-((S)-1-pyridin-2-yl-pyrrolidin-3-yloxy)-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-6-((S)-1-pyridin-2-yl-pyrrolidin-3-yloxy)-3,4-dihydro-2H-benzo[1,4]oxazine(prepared as described in example B1; 60 mg, 0.154 mmol),2-chloropyridine (CAS 109-09-1, 0.017 ml, 21.0 mg, 0.185 mmol), Xphos(8.81 mg, 0.018 mmol) and Cs₂CO₃ (125 mg, 0.385 mmol) in dioxane (1 ml)was degassed with argon, then Pd₂(dba)₃ (7.05 mg, 0.0077 mmol) wasadded. The reaction mixture was heated at 80° C. for 6 h, XPhos (8.81mg, 0.018 mmol) was added, the mixture was again degassed with argon andPd₂(dba)₃ (7.05 mg, 0.0077 mmol) was added. Stirring was continued overnight at 80° C. The mixture was filtered through celite and concentratedto give the title compound that was purified by NP-HPLC (column GraceGrom Saphir 65 Si, gradient heptane:EtOAc:MeOH 68:30:2 to 0:65:35 in 12min), yield 32 mg (45%).

HPLC Rt_(M1)=0.72 min; ESIMS: 467 [(M+H)⁺]

¹H NMR (400 MHz, CDCl₃): δ 8.32 (d, 1H), 8.14-8.12 (m, 1H), 7.49-7.39(m, 2H), 6.86 (d, 1H), 6.61 (d, 1H), 6.57-6.46 (m, 2H), 6.37 (d, 1H),4.92-4.85 (m, 1H), 4.26-4.24 (m, 2H), 3.76-3.74 (m, 2H), 3.71 (d, 2H),3.63-3.55 (m, 2H), 3.32 (s, 3H), 2.67 (s, 3H), 2.35-2.27 (m, 1H),2.26-2.15 (m, 1H).

Example N4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-6-((S)-1-pyrimidin-2-yl-pyrrolidin-3-yloxy)-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-6-((S)-1-pyridin-2-yl-pyrrolidin-3-yloxy)-3,4-dihydro-2H-benzo[1,4]oxazine(prepared as described in example B1; 60 mg, 0.154 mmol),2-chloropyrimidine (CAS 1722-12-9, 24.7 mg, 0.216 mmol) and DIPEA (0.054ml, 39.8 mg, 0.308 mmol) in ACN (1 ml) was heated at 140° C. for 30 minin a microwave reactor. The product was extracted with sat. aq. NaHCO₃soln. and EtOAc, filtered and concentrated to yield the title compoundthat was purified by prep. NP-HPLC (column Grace Grom Saphir 65 Si,gradient heptane:EtOAc:MeOH 68:30:2 to 0:65:35 in 12 min), yield 45 mg(63%)

HPLC Rt_(M1)=0.97 min; ESIMS: 468 [(M+H)⁺]

¹H NMR (400 MHz, CDCl₃): δ 8.37-8.26 (m, 3H), 7.44 (d, 1H), 6.87 (d,1H), 6.62 (d, 1H), 6.56-6.46 (m, 2H), 4.92-4.84 (m, 1H), 4.27-4.25 (m,2H), 3.90-3.63 (m, 6H), 3.33 (s, 3H), 2.69 (s, 3H), 2.37-2.13 (m, 2H).

Example O12-Methoxy-5-{2-methyl-6-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile

a) N-(5-Benzyloxy-2-hydroxy-phenyl)-2-chloro-propionamide

A solution of 2-chloro-propionic acid (CAS registry 598-78-7) (0.914 ml,7.53 mmol) in DMF (20 ml) was treated with Et₃N (1.259 ml, 9.03 mmol)and HATU (3.05 g, 8.03 mmol). The resulting solution was stirred at rtfor 30 min, then 2-amino-4-benzyloxy-phenol (CAS registry 102580-07-4)(1.08 g, 5.02 mmol) was added. The reaction mixture was stirred at rtfor 5 min, diluted with EtOAc and concentrated. The title compound wasobtained after flash chromatography on silica gel (cyclohexane/EtOAc,100:0 to 50:50) as orange solid (617 mg, 40% yield).

UPLC Rt_(M14)=1.32 min; ESIMS: 306 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 9.50 (d, 1H), 7.70 (br, s 1H), 7.45 (m,5H), 6.80 (d, 1H), 6.65 (dd, 1H), 5.00 (s, 2H), 2.65 (s, 3H).

b) 6-Benzyloxy-2-methyl-4H-benzo[1,4]oxazin-3-one

A dry solution of N-(5-benzyloxy-2-hydroxy-phenyl)-2-chloro-propionamide(617 mg, 2.0 mmol) in DMF (15 ml) was treated at 0° C. with sodiumhydride 95% (58.1 mg, 2.4 mmol). After stirring at rt for 1 h, thereaction mixture was diluted with DCM and washed with water. The organiclayer was dried over Na₂SO₄, filtered, concentrated and the titlecompound was obtained after flash chromatography on silica gel(cyclohexane/EtOAc, 100:0 to 80:20) as a white solid (146 mg, 27%yield).

UPLC Rt_(M14)=1.30 min; ESIMS: 270 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 10.80 (s, 1H), 7.45 (m, 5H), 6.85 (d, 1H),6.65 (m, 2H), 5.00 (s, 2H), 4.55 (q, 1H), 1.45 (d, 3H).

c) 6-Benzyloxy-2-methyl-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of 6-benzyloxy-2-methyl-4H-benzo[1,4]oxazin-3-one (146 mg,0.54 mmol) in THF (4 ml) was treated at 0° C. with BH₃*THF (1M in THF,0.813 ml, 0.813 mmol). After stirring at 35° C. for 1 h, the reactionmixture was cooled down to 0° C., quenched with water (0.5 ml) and anaqueous NaOH 4N soln. (0.5 ml) and then diluted with EtOAc. The organiclayer was dried over Na₂SO₄, filtered, concentrated and the titlecompound was obtained as a white solid (125 mg, 90% yield).

UPLC Rt_(M14)=1.40 min; ESIMS: 256 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.45 (m, 5H), 6.50 (d, 1H), 6.20 (d, 1H),6.10 (dd, 1H), 5.35 (s, 1H), 4.90 (s, 2H), 4.00 (m, 1H), 3.25 (m, 1H),2.90 (m, 1H), 1.35 (d, 3H).

d) 2-Methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol

A solution of 6-benzyloxy-2-methyl-3,4-dihydro-2H-benzo[1,4]oxazine (124mg, 0.486 mmol) in MeOH (10 ml) was treated at rt with ammonium formate(276 mg, 4.37 mmol) and Pd(OH)₂ (68.2 mg, 0.486 mmol). The reactionmixture was stirred at 60° C. for 15 min. After cooling to rt, thereaction mixture was filtered through hyflo, rinsed with DCM and MeOHand then the filtrates were concentrated. The title compound wasobtained after flash chromatography on silica gel (DCM/MeOH, 100:0 to90:10) as a brown solid. (69.4 mg, 87% yield).

UPLC Rt_(M14)=0.56 min; ESIMS: 166 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.50 (s, 1H), 6.45 (d, 1H), 6.00 (d, 1H),5.85 (dd, 1H), 5.25 (s, 1H), 4.00 (m, 1H), 3.25 (m, 1H), 2.90 (m, 1H),1.35 (d, 3H).

e)[(S)-3-(2-Methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy)-pyrrolidin-1-yl]-(tetrahydro-pyran-4-yl)-methanone

A dry solution of 2-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol (69 mg,0.42 mmol) and (R)-1-(tetrahydro-2H-pyran-4-carbonyl)pyrrolidin-3-ylmethanesulfonate (intermediate C1, 209 mg, 0.75 mmol) in DMF (1.4 ml)was treated with sodium hydride 60% in mineral oil (15.8 mg, 0.63 mmol)and the reaction mixture was stirred at 50° C. for 18 h. The reactionmixture was diluted with EtOAc and washed with sat. aq. NaHCO₃ soln. Theorganic layer was dried over Na₂SO₄, filtered, concentrated and thetitle compound was obtained after flash chromatography on silica gel(DCM/MeOH, 100:0 to 95:5) as a red orange sticky solid (128 mg, 88%yield).

UPLC Rt_(M14)=1.01 min; ESIMS: 347 [(M+H)⁺]

¹H NMR (400 MHz, DMSO-d₆): δ 6.50 (d, 1H), 6.25 (d, 1H), 6.00 (d, 1H),6.05 (m, 1H), 5.65 (m, 2H), 5.35 (d, 2H), 4.45 (d, 2H), 3.00-4.00 (m,6H), 2.00-2.40 (m, 4H), 1.5 (m, 4H)

f)2-Methoxy-5-{2-methyl-6-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile

A mixture of(S)-(3-(2-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yloxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone(128 mg, 0.369 mmol), 5-bromo-2-methoxynicotinonitrile (CAS registry941294-54-8, IA12), (94 mg, 0.443 mmol), XPhos (8.81 mg, 0.018 mmol),NaOtBu (53.3 mg, 0.554 mmol) and Pd₂(dba)₃ (16.92 mg, 0.018 mmol) intoluene (2.5 ml) was degassed with argon. The reaction mixture wasstirred at 80° C. for 20 min. After cooling to rt, the reaction mixturewas filtered through hyflo, rinsed with EtOAc and the filtrates werewashed with sat. aq. NaHCO₃ soln. The organic layer was dried overNa₂SO₄, filtered and concentrated. The title compound was obtained afterprep. RP-HPLC (SunFire C18 column OBD 5 mm 30×100 mm, gradient 32% to67% ACN in 15 min). The fractions were lyophilized and filtered over aPL-HCO₃ MP SPE cartridge to give a brown solid (39.1 mg, 22% yield).

UPLC Rt_(M14)=1.10 min; ESIMS: 479 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆, 394 K): δ 8.40-8.30 (m, 1H), 8.10-8.00 (m,1H), 6.75 (d, 1H), 6.35 (m, 1H), 6.15 (m, 1H), 4.75 (m, 1H), 4.00 (s,3H), 3.85 (m, 1H), 3.75-3.00 (m, 5H), 2.65 (m, 1H), 2.00 (m, 1H), 1.65(m, 2H), 1.44 (d, 3H).

Examples O2 to O3

The compounds listed in Table 10 were prepared by chromatographicdiastereomer separation.

TABLE 10 HPLC Rt MS Compound/ [min] [m/z; Example Reaction Conditions(method) (M + 1)⁺] O2

  Peak 1 diastereomer separation2-Methoxy-5-{(S)-2-methyl-6-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile Buchwald aminationcondition: CA6 Amide bond condition: CB6 Side chain introductioncondition: CC2 Precursors used: IO, IA12, CAS 104706-47-0/ acyl chloride40191-32-0 Chromatographic diasteroemer separation: CD11 19.89 (M15) O3

  Peak 2 diastereomer separation2-Methoxy-5-{(R)-2-methyl-6-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile Buchwald aminationcondition: CA6 Amide bond condition: CB6 Side chain introductioncondition: CC2 Precursors used: IO, IA12, CAS 104706-47-0/ Acyl chloride40191-32-0 Chromatographic diastereomer separation: CD11 27.33 (M15)

Example P2-Methoxy-5-{6-[(S)-1-(1-methyl-piperidin-4-ylmethyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile

a) (S)-tert-butyl4-((3-(4-(5-cyano-6-methoxypyridin-3-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yloxy)pyrrolidin-1-yl)methyl)piperidine-1-carboxylate

A solution of2-methoxy-5-[6-((S)-pyrrolidin-3-yloxy)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-nicotinonitrile(see analogue B1, c), 95 mg, 0.270 mmol) in DCE (4.5 ml) was treatedwith tert-butyl 4-formylpiperidine-1-carboxylate (60 mg, 0.281 mmol).After stirring for 2 d at rt, the reaction mixture was diluted with DCMand sat. aq. NaHCO₃ soln. The organic layer was dried over Na₂SO₄,filtered, concentrated and the title compound was obtained after flashchromatography on silica gel (cyclohexane/EtOAc, 100:0 to 0:100), yield66 mg, (40%).

UPLC Rt_(M1)=1.66 min; ESIMS: 550 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.35 (d, 1H), 7.75 (d, 1H), 7.35 (s, 1H),7.85 (d, 1H), 6.35 (dd, 1H), 6.10 (d, 1H), 4.65 (m, 1H), 4.45 (m, 2H),3.60 (m, 2H), 2.75-2.15 (m, 4H), 1.50 (s, 9H).

b)(S)-2-methoxy-5-(6-(1-(piperidin-4-ylmethyl)pyrrolidin-3-yloxy)-2H-benzo[b][1,4]oxazin-4(3H)-yl)nicotinonitrile

A solution of (S)-tert-butyl4-((3-(4-(5-cyano-6-methoxypyridin-3-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yloxy)pyrrolidin-1-yl)methyl)piperidine-1-carboxylate)(66 mg, 0.120 mmol) in DCM (2 ml) was treated with TFA (0.093 ml, 1.20mmol). The reaction mixture was stirred at rt for 17 h, then quenchedwith sat. aq. Na₂CO₃ soln. and extracted with DCM. The organic layer wasdried over Na₂SO₄, filtered and concentrated under reduced pressure toafford the title product (36 mg, 67% yield).

UPLC Rt_(M1)=1.03 min; ESIMS: 450 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.35 (d, 1H), 7.80 (d, 1H), 7.25 (s, 1H),6.80 (d, 1H), 6.25 (m, 1H), 6.15 (d, 1H), 4.65 (m, 1H), 4.45 (m, 2H),3.60 (m, 2H), 2.75-2.15 (m, 4H).

c)(S)-2-methoxy-5-(6-(1-((1-methylpiperidin-4-yl)methyl)pyrrolidin-3-yloxy)-2H-benzo[b][1,4]oxazin-4(3H)-yl)nicotinonitrile

A solution of(S)-2-methoxy-5-(6-(1-(piperidin-4-ylmethyl)pyrrolidin-3-yloxy)-2H-benzo[b][1,4]oxazin-4(3H)-yl)nicotinonitrile(36 mg, 0.080 mmol) in DCE (2 ml) was treated with a 37% aq.formaldehyde soln. (8.94 μl, 0.120 mmol). The solution was stirred underargon at rt for 15 min, then NaBH₃CN (50.9 mg, 0.240 mmol) was added.The resulting mixture was stirred at rt for 30 min, diluted with DCM andsat. aq. NaHCO₃ soln. The organic layer was dried over Na₂SO₄ andconcentrated. The title compound was obtained after purification byprep. RP-HPLC (column SunFire C18, gradient 15-50% ACN in 15 min). Thefractions were extracted with DCM/sat. aq. NaHCO₃ soln, dried overNa₂SO₄, concentrated and lyophilized to give the title compound (18 mg,48% yield).

UPLC Rt_(M1)=1.04 min; ESIMS: 464 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.35 (d, 1H), 7.85 (d, 1H), 7.45 (s, 1H),6.75 (d, 1H), 6.35 (dd, 1H), 6.15 (d, 1H), 4.65 (m, 1H), 4.45 (m, 2H),3.25 (m, 2H), 2.75-2.15 (m, 4H), 2.65 (s, 3H), 1.95 (m, 2H), 1.65 (m,2H).

Example Q{(S)-3-[4-(6-Methoxy-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

a) 6-(tert-Butyl-dimethyl-silanyloxy)-4H-benzo[1,4]oxazin-3-one

A solution of 6-hydroxy-2H-benzo[b][1,4]oxazin-3(4H)-one (CAS registry53412-38-7) (1076 mg, 6.52 mmol) in DMF (8 ml) was treated at rt withTBDMSCl (1080 mg, 7.17 mmol) and imidazole (532 mg, 7.82 mmol). Afterstirring for 18 h at rt, the reaction mixture was diluted with DCM andwashed with water. The organic layer was dried over Na₂SO₄, filtered,concentrated and the title compound was obtained after flashchromatography on silica gel (cyclohexane/EtOAc, 100:0 to 50:50) as awhite solid (1.18 g, 65% yield).

UPLC Rt_(M2)=1.91 min; ESIMS: 280 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.85 (br, s 1H), 7.35 (s, 1H), 6.85 (d, 1H),6.45 (m, 1H), 6.30 (d, 1H), 4.50 (s, 2H), 1.00 (s, 9H), 0.25 (s, 6H).

b) 3,3-Dideutero-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol

A solution of6-(tert-butyl-dimethyl-silanyloxy)-4H-benzo[1,4]oxazin-3-one (8.34 g,29.8 mmol) in THF (100 ml) was treated at 00° C. with lithium aluminiumdeuteride (2.26 g, 59.7 mmol). After stirring for 18 h at rt thereaction mixture was added to a cold aqueous 1 M Rochelle's salt soln.and was extracted with EtOAc. The organic layer was dried over Na₂SO₄,filtered, concentrated and the title compound was obtained after flashchromatography on silica gel (cyclohexane/EtOAc, 100:0 to 0:100) as awhite solid (1.40 g, 31% yield).

UPLC Rt_(M9)=0.69 min; ESIMS: 154 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.50 (s 1H), 6.45 (d, 1H), 6.00 (d, 1H),5.85 (m, 1H), 5.15 (s, 1H), 4.00 (s, 2H).

c)(S)-3-(3,3-Dideutero-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

A dry solution of 3,3-dideutero-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol(1.44 g, 9.40 mmol) and(R)-3-methanesulfonyloxy-pyrrolidine-1-carboxylic acid tert-butyl ester(CAS registry 127423-61-4) (5.49 g, 20.68 mmol) in DMF (10 ml) wastreated with sodium hydride 60% in mineral oil (0.752 g, 18.80 mmol) andthe reaction mixture was stirred at rt for 2 d. The reaction mixture wasdiluted with EtOAc and washed with sat. aq. NaHCO₃ soln. The organiclayer was dried over Na₂SO₄, filtered, concentrated and purified byflash chromatography on silica gel (cyclohexane/EtOAc, 100:0 to 0:100),to yield 4.12 g, (quantitative yield) of the title compound.

UPLC Rt_(M) i=1.07 min; ESIMS: 323 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 6.65 (m, 1H), 6.15 (m, 2H), 5.35 (m, 1H),4.85 (m, 2H), 3.50 (m, 4H), 2.15 (m, 2H), 1.50 (s, 9H).

d)(S)-3-[4-(6-Methoxy-5-methyl-pyridin-3-yl)-3,3-dideutero-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

This example was prepared in analogy to Example G1, e).

UPLC Rt_(M1)=2.00 min; ESIMS: 444 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.95 (br.s, 1H), 7.45 (br.s, 1H), 6.80 (m,1H), 6.25 (m, 1H), 4.75 (m, 1H), 4.35 (s, 2H), 4.00 (s, 3H), 3.50 (m,4H), 2.25 (m, 2H), 1.50 (s, 9H).

e)4-(6-Methoxy-5-methyl-pyridin-3-yl)-3,3-dideutero-6-((S)-pyrrolidin-3-yloxy)-3,4-dihydro-2H-benzo-[1,4]oxazine

This example was prepared in analogy to Example G1, f).

UPLC Rt_(M1)=1.26 min; ESIMS: 342 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.95 (br.s, 1H), 7.45 (br.s, 1H), 6.80 (m,1H), 6.25 (m, 1H), 4.75 (m, 1H), 4.35 (s, 2H), 4.00 (s, 3H), 3.15 (m,2H), 2.75 (m, 2H), 2.25 (m, 2H).

f){(S)-3-[4-(6-Methoxy-5-methyl-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

This example was prepared in analogy to Example B1, d). UPLCRt_(M1)=1.65 min; ESIMS: 456 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃, 298 K): δ 8.45 (m, 1H), 8.29 (s, 1H), 7.21 (t,1H), 6.20 (t, 1H), 6.10 (s, 1H), 5.00 (d, 1H), 4.37 (t, 2H), 4.00 (m,3H), 3.39-3.74 (m, 4H), 2.50 (m, 2H), 2.35 (s, 3H), 1.09-2.10 (m, 5H).

Example R5-{6-[(S)-1-(4-Hydroxy-cyclohexanecarbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-2-methoxy-nicotinonitrile

This example was prepared in analogy to Example J, starting from2-methoxy-5-{6-[(S)-1-((S)-pyrrolidine-3-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile.

UPLC Rt_(M14)=0.91 min; ESIMS: 478 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d6): δ 8.43 (d, 1H), 8.28 (d, 1H), 6.73 (m, 1H),6.34 (m, 1H), 6.08 (dd, 1H), 4.76 (d, 1H), 4.51 (m, 1H), 4.22 (s, 2H),4.00 (s, 3H), 3.20-3.71 (m, 9H), 1.09-2.10 (m, 10H).

Example S2-Methoxy-5-{6-[(S)-1-(2-pyridin-4-yl-acetyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile

This example was prepared in analogy to Example J, starting from2-methoxy-5-{6-[(S)-1-((S)-pyrrolidine-3-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile.

UPLC Rt_(M14)=0.82 min; ESIMS: 471 [(M+H)⁺]

¹H NMR (400 MHz, DMSO-d6): δ 8.45 (m, 3H), 8.29 (s, 1H), 7.21 (m, 2H),6.74 (m, 1H), 6.33 (m, 1H), 6.10 (m, 1H), 4.87 (d, 1H), 4.22 (s, 2H),4.00 (s, 3H), 3.39-3.74 (m, 8H), 1.09-2.10 (m, 2H).

Example T{(S)-3-[4-(5-Amino-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanonea) 5-[6-((S)-1-tert-Butoxycarbonyl-pyrrolidin-3-yloxy)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-2-methoxy-nicotinic acid methyl ester

Under argon, K₃PO₄ (815 mg, 2.00 mmol) andbis-(t-butylphosphine)palladium (29.4 mg. 0.06 mmol) were added to asolution of(S)-3-(3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (prepared as described in step a) example B) (615mg, 1.92 mmol) and 5-bromo-2-methoxy-nicotinic acid methyl ester (IA 22,CAS registry 122433-41-4) (614 mg, 1.30 mmol) in toluene (6 ml). Thereaction mixture was degassed with argon for 15 min, then stirred at110° C. for 18 h, diluted with EtOAc and washed with a sat. aq. NaHCO₃soln. The organic layer was dried over MgSO₄ and concentrated to affordthe crude title compound that was purified by flash chromatography onsilica gel (heptane/EtOAc 90:10 to 0:100) to give a yellow gum (474 mg,51% yield).

UPLC Rt_(M2)=1.36 min; ESIMS: 486 [(M+H)⁺].

b)5-[6-((S)-1-tert-Butoxycarbonyl-pyrrolidin-3-yloxy)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-2-methoxy-nicotinicacid

A solution of5-[6-((S)-1-tert-butoxycarbonyl-pyrrolidin-3-yloxy)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-2-methoxy-nicotinicacid methyl ester (483 mg, 0.99 mmol) in dioxane (5 ml) was treated witha solution of sodium hydroxide pellets (119 mg, 2.98 mmol) in water (2ml). The solution was stirred at 80° C. for 1 h. The reaction mixturewas acidified to pH3 with aq. 1N HCl soln. and extracted with EtOAc. Thecombined organic phases were dried over MgSO₄ and concentrated to affordthe title compound after flash chromatography on silica gel(heptane/EtOAc 100:0 to 0:100 then EtOAc/MeOH 90:10 to 80:20) as a solid(370 mg, 79% yield).

UPLC Rt_(M6)=1.79 min; ESIMS: 372 [(M+H-100)⁺]

¹H NMR (400 MHz, CDCl₃): δ 8.22-8.48 (m, 2H), 6.82 (d, 1H), 6.33 (m,1H), 6.21 (d, 1H), 4.60-4.76 (m, 1H), 4.27-4.41 (m, 2H), 4.15-4.27 (m,3H), 3.62-3.77 (m, 2H), 3.31-3.58 (m, 5H), 1.85-2.19 (m, 2H), 1.34-1.56(m, 9H)

c)(S)-3-[4-(5-tert-Butoxycarbonylamino-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylic acid tert-butylester

A solution of5-[6-((S)-1-tert-butoxycarbonyl-pyrrolidin-3-yloxy)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-2-methoxy-nicotinicacid (370 mg, 0.78 mmol) and Et₃N (0.28 ml, 1.96 mmol) in tBuOH (5 ml)was treated with DPPA (CAS registry 26386-88-9) (0.17 ml, 0.78 mmol) andstirred at 100° C. for 6 h. DCM and sat. aq. NaHCO₃ soln. were added,the organic layer was separated by elution through a separating phasecartridge and concentrated to afford the title compound after flashchromatography on silica gel (heptane/EtOAc 100:0 to 50:50) as a pinkgum (114 mg, 24%).

UPLC Rt_(M2)=1.36 min; ESIMS: 486 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.42 (br s, 1H), 7.75 (d, 1H), 7.02 (s, 1H),6.78 (d, 1H), 6.15-6.41 (m, 1H), 4.71 (br s, 1H), 4.23-4.39 (m, 2H),4.09-4.22 (m, 3H), 3.62-3.75 (m, 2H), 3.31-3.58 (m, 4H), 1.86-2.26 (m,2H), 1.40-1.60 (m, 18H).

d)2-Methoxy-5-[6-((S)-pyrrolidin-3-yloxy)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-pyridin-3-ylamine

A solution of(S)-3-[4-(5-tert-butoxycarbonylamino-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (130 mg, 0.24 mmol) in DCM (2 ml) was treated withTFA (CAS registry 76-05-1) and stirred at rt for 1 h. The reactionmixture was concentrated to afford the title compound after elution froma 2 g Isolute SCX-2 cartridge (eluent MeOH, then 2M NH₃/MeOH) as ayellow gum (88 mg, quant. crude).

UPLC Rt_(M2)=1.25 min; ESIMS: 343 [(M+H)⁺].

e){(S)-3-[4-(5-Amino-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone

A solution of 1-methyl-1H-imidazole-4-carboxylic acid (CAS registry41716-18-1) (36.0 mg, 0.26 mmol) and Et₃N (0.11 ml, 0.78 mmol) in DMF (1ml) was treated with HBTU (CAS registry 94790-37) (107 mg, 0.28 mmol).After stirring at rt for 20 min, the reaction mixture was cooled down to5° C. and a solution of2-methoxy-5-[6-((S)-pyrrolidin-3-yloxy)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-pyridin-3-ylamine(88 mg, 0.26 mmol) in DMF (3 ml) was added. The reaction mixture wasstirred at rt for 1 h, concentrated and the residue was taken up in DCM(10 ml), washed with a sat. aq. NaHCO₃ soln. (5 ml), the organic layerwas separated by elution through a separating phase cartridge andconcentrated. The residue was purified by flash chromatography on silicagel (heptane/EtOAc 100:0 to 0:100), the combined fractions wereconcentrated, dissolved in tBuOH/H₂O and lyophilized to afford the titlecompound as a colourless solid (21 mg, 37% yield).

UPLC Rt_(M2)=0.85 min; ESIMS: 451 [(M+H)⁺].

¹H NMR (400 MHz, CD₃OD): δ 7.64 (s, 1H), 7.62 (s, 1H), 7.65 m, 1H), 6.93(m, 1H), 6.70 (m, 1H), 6.20-6.34 (m, 1H), 6.15 (m, 1H), 4.81 (m, 1H),4.19-4.31 (m, 2H), 3.90-4.05 (m, 4H), 3.55-3.83 (m, 8H), 2.04-2.28 (m,2H).

Example UN-(2-Methoxy-5-{6-[(S)-1-(1-methyl-1H-imidazole-4-carbonyl)-pyrrolidin-3-yloxy]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-pyridin-3-yl)-methanesulfonamide

A solution of{(S)-3-[4-(5-Amino-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]-oxazinyloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone(22.9 mg, 0.05 mmol) in pyridine (1 ml) was treated with methanesulfonylchloride (CAS registry 124-63-0) (0.08 ml, 0.97 mmol) and stirred at 50°C. for 18 h. The reaction mixture was diluted with DCM and H₂O, theorganic layer was separated by elution through a separating phasecartridge and concentrated. The residue was purified by flashchromatography on silica gel (heptane/EtOAc 100:0 to 0:100 thenEtOAc/MeOH 90:10 to 80:20), the combined fractions were concentrated,dissolved in tBuOH/H₂O and lyophilized to afford the title compound as acolourless solid (14 mg, 49%).

UPLC Rt_(M2)=1.39 min; ESIMS: 529 [(M+H)⁺].

¹H NMR (400 MHz, CD₃OD): δ 7.87 (d, 1H), 7.41-7.76 (m, 3H), 6.73 (m,1H), 6.02-6.44 (m, 2H), 4.14-4.39 (m, 2H), 3.87-4.12 (m, 5H), 3.55-3.84(m, 8H), 2.85-3.08 (m, 3H), 1.75-2.40 (m, 2H).

Example V(1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[4-(6-methoxy-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-methanone

a) 6-Methoxy-5-methyl-4H-benzo[1,4]oxazin-3-one

A solution of 6-bromo-5-methyl-4H-benzo[1,4]oxazin-3-one (CAS registry1154740-47-2) (1000 mg, 4.13 mmol) and CuI (79 mg, 0.41 mmol) in NaOMe30% in MeOH (8.1 ml) was stirred at 130° C. for 5 h. The orange/brownmixture was cooled to rt, diluted with EtOAc and washed with a sat. aq.NaHCO₃ soln. The combined organic layers were dried over MgSO₄, andconcentrated to afford an orange solid. Trituration with cyclohexaneafforded the title compound as a pink solid (647 mg, 70% yield).

HPLC Rt_(M1)=0.73 min.

¹H NMR (400 MHz, DMSO-d₆): δ 10.21 (s, 1H), 6.76 (d, 1H), 6.53 (d, 1H),4.42 (s, 2H), 3.71 (s, 3H), 2.05 (s, 3H).

b) 6-Hydroxy-5-methyl-4H-benzo[1,4]oxazin-3-one

A suspension of 6-methoxy-5-methyl-4H-benzo[1,4]oxazin-3-one (647 mg,3.35 mmol) in DCM (30 ml) was treated under argon at rt with BBr₃ (3.16ml, 33.5 mmol) and stirred for 18 h at rt. The reaction mixture wasquenched by dropwise addition of MeOH at 0° C. until obtention of aclear solution. After removal of the solvents, the residue was pouredonto ice/sat. aq. NaHCO₃ soln. and extracted with EtOAc. The organiclayer was dried over MgSO₄ and concentrated to afford the title compoundas a brown solid (647 mg, crude), which was used in the next stepwithout further purification.

HPLC Rt_(M1)=0.49 min.

¹H NMR (400 MHz, DMSO-d₆): δ 10.11 (s, 1H), 6.43 (d, 1H), 5.98 (d, 1H),4.57 (s, 2H), 1.89 (s, 3H).

c) 5-Methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol

A solution of 6-hydroxy-5-methyl-4H-benzo[1,4]oxazin-3-one (647 mg, 3.61mmol) in THF (20 ml) was treated under argon at 0° C. with BH₃*THF (1Min THF, 10.83 ml, 10.83 mmol) and stirred for 18 h at rt. MeOH was addedand the solution was stirred at rt for 1 h, concentrated to afford aresidue which was dissolved in THF (20 ml), treated with BH₃*THF (1M inTHF, 10.83 ml, 10.83 mmol) and stirred at rt for 18 h. MeOH was added,the solution was stirred at rt for 4 h, concentrated to dryness toafford the title compound as a brown solid (600 mg, crude), which wasused in the next step without further purification.

HPLC Rt_(M1)=0.49 min; ESIMS: 166 [(M+H)⁺].

d)(S)-3-(5-Methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

A solution of triphenylphosphine (1.33 g, 5.09 mmol) in THF (10 ml) wastreated with DEAD (0.8 ml, 5.09 mmol) followed by(R)-3-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (CASregistry 127423-61-4) (1 g, 5.45 mmol) and5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-01 (600 mg, 3.63 mmol). Theresulting red/brown solution was stirred at 70° C. for 18 h. The brownmixture was cooled down, diluted with EtOAc and washed with sat. aq.NaHCO₃ soln. The combined organic layers were dried over MgSO₄, filteredand concentrated to afford a brown oil. The crude product was threetimes purified by flash chromatography on silica gel (cyclohexane/EtOAc90:10 to 40:60) to afford the title compound as a colourless oil (130mg, 11% yield)

HPLC Rt_(M1)=1.09 min; ESIMS: 335 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 6.44 (d, 1H), 6.22-6.04 (m, 1H), 5.24 (brs, 1H), 4.75 (br s, 1H), 4.07-3.93 (m, 2H), 3.45-3.33 (m, 3H), 3.28 (d,7H), 2.00 (d, 2H), 1.83 (d, 3H), 1.46-1.32 (m, 9H).

e)(S)-3-[4-(6-Methoxy-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

A solution of(S)-3-(5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (120 mg, 0.36 mmol),5-bromo-2-methoxy-3-methylpyridine (CAS registry 760207-87-2) (145 mg,0.72 mmol), NaOtBu (103 mg, 1.08 mmol), RuPhos (CAS registry787618-22-8) (8 mg, 0.02 mmol) and [RuPhos]palladacycle (CAS registry787618-22-8) (15 mg, 0.02 mmol) in dioxane (2 ml) was stirred at 100° C.for 18 h. The orange/brown mixture was cooled, diluted with EtOAc andwashed with water. The combined organic layers were dried over MgSO₄,filtered and concentrated to afford a brown oil. The crude product wasthree times purified by flash chromatography on silica gel(cyclohexane/EtOAc 95:05 to 60:40) to afford the title compound as ayellow oil (97 mg, 60% yield)

HPLC Rt_(M1)=1.37 min; ESIMS: 456 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.40 (d, 1H), 7.28-7.07 (m, 1H), 6.74 (s,2H), 4.84 (br s, 1H), 3.96 (br s, 2H), 3.80 (s, 2H), 3.57 (br s, 2H),3.44-3.19 (m, 10H), 2.08 (s, 3H), 2.05-1.92 (m, 2H), 1.60 (d, 3H), 1.34(d, 9H).

f) (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[4-(6-methoxy-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-methanone

A solution of(S)-3-[4-(6-methoxy-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (97 mg, 0.21 mmol) in DCM (3 ml) was treated underargon at rt with TFA (0.16 ml, 2.13 mmol) and stirred for 6 h. Thereaction mixture was quenched with sat. aq. NaHCO₃ soln. and the organicsolution was separated through a phase separating cartridge affording ayellow solution. 1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-carboxylicacid (CAS registry 64096-87-3) (49 mg, 0.28 mmol), Et₃N (0.09 ml, 0.64mmol), EDC (62 mg, 0.32 mmol), HOBT (49 mg, 0.32 mmol) were added to theyellow solution and stirred at rt for 18 h. The reaction mixture wasquenched with sat. aq. NaHCO₃ soln. and the organic layer was separatedby passing through a phase separating cartridge, then concentrated andpurified by prep. RP-HPLC (column SunFire C18 OBD 5 mm 30×100 mm,Solvent A: H₂O (0.1% TFA) Solvent B: CH₃CN (0.1% TFA) afforded the titlecompound as white solid (76 mg, 70% yield)

HPLC Rt_(M1)=0.98 min; ESIMS: 516 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆, 375K): δ 7.44 (br s, 1H), 7.18 (br s, 1H),6.75 (s, 2H), 4.88 (br s, 1H), 4.02 (t, 2H), 3.86 (s, 3H), 3.81-3.33 (m,6H), 3.24-3.06 (m, 4H), 2.83 (br s, 1H), 1.66 (s, 3H). Rotamers.

Example W{(S)-3-[4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanoneLe

a) 6-Bromo-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of 6-bromo-5-methyl-4H-benzo[1,4]oxazin-3-one (CAS registry1154740-47-2) (425 mg, 1.56 mmol) in THF (9 ml) was treated under argonat 0° C. with BH₃*THF (1M in THF, 4.7 ml, 4.69 mmol) and stirred for 18h at rt. BH₃.THF 1M (2 ml) was added and stirring was continued foranother 24 h. The reaction mixture was concentrated and purified byflash chromatography on silica gel (cyclohexane/EtOAc 100:0 to 80:20) toafford the title compound as an orange solid (324 mg, 86% yield)

HPLC Rt_(M1)=1.04 min; ESIMS: 228, 230 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 6.68 (d, 1H), 6.48 (d, 1H), 5.54 (br s,1H), 4.04 (t, 2H), 3.36-3.26 (m, 2H), 2.11 (s, 3H).

b)6-Bromo-4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of 6-bromo-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazine (324 mg,1.42 mmol), Intermediate IA1 (391 mg, 1.56 mmol), Cs₂CO₃ (1018 mg, 3.13mmol), BINAP (CAS registry 98327-87-8) (44 mg, 0.07 mmol), Pd(OAc)₂ (CASregistry 3375-31-3) (32 mg, 0.14 mmol) in toluene (13 ml) was stirred at100° C. for 18 h. Catalyst and ligand were reloaded and stirring wascontinued for another 24 h at 100° c. The reaction mixture was cooled tort, diluted with EtOAc and washed with water. Concentration of theorganic layer and purification by flash chromatography on silica gel(cyclohexane/EtOAc 97:03 to 40:60) afforded the title compound as anorange solid (345 mg, 58% yield).

HPLC Rt_(M) i=1.13 min; ESIMS: 397,399 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.99 (br. s, 1H), 7.37 (d, 1H), 7.24 (br.s, 1H), 6.83 (d, 1H), 4.13 (t, 2H), 3.97-3.86 (m, 2H), 3.30 (s, 3H),2.52 (s, 3H), 1.93 (s, 3H).

c)Benzyl-[4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yl]-amine

A solution of6-bromo-4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazine(324 mg, 0.82 mmol), benzylamine (350 mg, 3.26 mmol), NaOtBu (157 mg,1.63 mmol), RuPhos (CAS registry 787618-22-8) (30 mg, 0.06 mmol) and[BrettPhos]palladacycle (CAS registry 1148148-01-9) (52 mg, 0.06 mmol)in dioxane (16 ml) was stirred at 80° C. for 0.5 h. Filtration,concentration of the filtrate and purification by flash chromatographyon silica gel (cyclohexane/EtOAc 88:12 to 35:65) afforded the titlecompound as a yellow oil (228 mg, 66% yield).

HPLC Rt_(M1)=1.13 min; ESIMS: 424 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.94 (br s, 1H), 7.40-7.33 (m, 2H), 7.30(t, 2H), 7.19 (t, 2H), 6.58 (d, 1H), 6.30 (d, 1H), 5.25 (t, 1H), 4.30(d, 2H), 4.04-3.97 (m, 2H), 3.90 (d, 2H), 3.29 (s, 3H), 2.51 (br s, 3H),1.76 (s, 3H).

d)4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylamine

A solution ofbenzyl-[4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yl]-amine(228 mg, 0.53 mmol), acetic acid (0.21 ml, 3.70 mmol), Pd/C in MeOH/THF(2.5/2.5 ml) was hydrogenated with H₂ at rt for 65 h. Filtration andconcentration of the filtrate afforded the title compound as a green oil(200 mg, crude, including remaining AcOH).

HPLC Rt_(M1)=0.64 min; ESIMS: 334 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.96 (br s, 1H), 7.17 (br s, 1H), 6.62-6.54(d, 1H), 6.54-6.45 (d, 1H), 4.48 (br s, 2H), 4.01 (t, 2H), 3.88 (br s,2H), 3.28 (s, 3H), 1.88 (d, 3H), 1.63 (s, 3H).

e)4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol

A solution of4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylamine(200 mg, 0.60 mmol) in water (3.5 ml) and H₂SO₄ (0.32 ml) was addeddropwise to a solution of sodium nitrite (49.7 mg, 0.72 mmol) in water(10 ml) at 0° C. The mixture was stirred at rt for 3 d. The reactionmixture was filtered and the filtrate was quenched with sat. aq. NaHCO₃soln. and extracted with EtOAc. The organic layer was dried over MgSO₄,filtered and concentrated to afford a brown oil. Purification by flashchromatography on silica gel (DCM/MeOH 88:12 to 80:20) afforded thetitle compound as a brown oil (50 mg, 25% yield).

HPLC Rt_(M1)=0.78 min; ESIMS: 335 [(M+H)⁺].

f)(1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[4-(6-methoxy-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-methanone

A solution of triphenylphosphine (55 mg, 0.21 mmol) in THF (2.5 ml) wastreated with DEAD (0.03 ml, 0.21 mmol), followed by(R)-3-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (CASregistry 127423-61-4) (33 mg, 0.18 mmol) and4-(6-methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol(50 mg, 0.15 mmol). The resulting red/brown solution was stirred at 70°C. for 18 h. The reaction mixture was cooled down to rt, diluted withEtOAc and washed with sat. aq. NaHCO₃ soln. The organic layer was driedover MgSO₄. concentrated and purified by flash chromatography on silicagel (cyclohexane/EtOAc 90:10 to 40:60) to afford the title compound asan orange solid (44 mg, 58% yield) HPLC Rt_(M1)=1.16 min; ESIMS: 504[(M+H)⁺].

g){(S)-3-[4-(6-Methanesulfonyl-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

A solution of(1,1-dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[4-(6-methoxy-5-methyl-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-methanone(44 mg, 0.09 mmol) in DCM (4 ml) was treated under argon at rt with TFA(0.07 ml, 0.17 mmol) and stirred for 18 h. The reaction mixture wasquenched with sat. aq. NaHCO₃ soln. and the organic solution wasseparated through a phase separating cartridge, concentrated andpurified by flash chromatography on silica gel (DCM/MeOH 100:0 to90:10). The obtained product was dissolved in DCM (4 ml) and Et₃N wasadded. Tetrahydro-pyran-4-carbonyl chloride (CAS registry 40191-32-0)(15 mg, 0.10 mmol) was added to the reaction mixture at 0° C. and theresulting orange solution was stirred at rt for 4 h. The reactionmixture was quenched with sat. aq. NaHCO₃ soln. and the organic layerwas separated by elution through a phase separating cartridge,concentrated and purified by SFC (column NH₂ (250×30 mm (l×w), 60 A, 5μm, Princeton, gradient of methanol in supercritical CO₂) to afford thetitle compound as a yellow oil (15 mg, 32% yield)

HPLC Rt_(M1)=0.88 min; ESIMS: 516 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆, 375K): δ 7.98 (br s, 1H), 7.19 (br s, 1H),6.92-6.85 (m, 1H), 6.85-6.77 (m, 1H), 4.95 (br s, 1H), 4.12 (t, 2H),3.92 (t, 2H), 3.88 (br s, 2H), 3.61 (br s, 3H), 3.38 (td, 2H), 3.27 (s,3H), 2.68 (d, 1H), 2.56 (s, 3H), 2.17 (br s, 2H), 1.73 (s, 3H), 1.58 (brs, 4H). Rotamers.

Example X{(S)-3-[4-(5-Difluoromethyl-6-methoxy-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-methanone

a) 6-Bromo-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of 6-bromo-5-methyl-4H-benzo[1,4]oxazin-3-one (CAS registry1154740-47-2) (2.8 g, 11.56 mmol) in THF (50 ml) was treated under argonwith BH₃*THF (1M in THF, 34.7 ml, 34.70 mmol) and heated under refluxfor 2 h. MeOH was added and the solution was stirred at rt for 1 h,concentrated and purified by flash chromatography on silica gel(cyclohexane/EtOAc 100:0 to 80:20) to afford the title compound as anorange solid (1.8 g, 68% yield).

HPLC Rt_(M1)=1.04 min; ESIMS: 228, 230 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 6.68 (d, 1H), 6.48 (d, 1H), 5.54 (br s,1H), 4.04 (t, 2H), 3.36-3.26 (m, 2H), 2.11 (s, 3H).

b)6-Bromo-4-(5-difluoromethyl-6-methoxy-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of 6-bromo-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazine (500 mg,2.19 mmol), Intermediate IA6 (574 mg, 2.41 mmol), NaOtBu (421 mg, 4.38mmol), BrettPhos (CAS registry 1070663-78-3) (59 mg, 0.11 mmol) and[BrettPhos]palladacycle (CAS registry 1148148-01-9) (88 mg, 0.11 mmol)in dioxane (11 ml) was stirred at 100° C. for 18 h. Catalyst and ligandwere reloaded and stirring was continued at 100° C. for 48 h. Thereaction mixture was cooled down to rt, diluted with EtOAc and washedwith sat. aq. NaHCO₃ soln. The organic layer was dried over MgSO₄,concentrated to afford a brown oil. Purification by flash chromatographyon silica gel (cyclohexane/EtOAc 100:0 to 80:20) afforded the titlecompound as a brown oil (150 mg, 18% yield).

HPLC Rt_(M1)=1.34 min; ESIMS: 385, 387 [(M+H)⁺].

c)4-(5-Difluoromethyl-6-methoxy-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol

A mixture of6-bromo-4-(5-difluoromethyl-6-methoxy-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazine(150 mg, 0.39 mmol), KOH (65 mg, 1.17 mmol) in water (0.33 ml),tetramethyl-t-butyl-XPhos (CAS registry 857356-94-6) (18.72 mg, 0.04mmol) and Pd₂(dba)₃ (17.83 mg, 0.02 mmol) in dioxane (2 ml) was degassedwith nitrogen and heated at 100° C. for 18 h. Filtration, concentrationand purification by flash chromatography on silica gel(cyclohexane/EtOAc 100:0 to 70:30) afforded the title compound as anorange oil (65 mg, 52% yield).

HPLC Rt_(M1)=1.01 min; ESIMS: 323 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.83 (s, 1H), 7.89 (d, 1H), 7.36 (d, 1H),7.00 (t, 1H), 6.61 (d, 1H), 6.55 (d, 1H), 3.93 (t, 2H), 3.88 (s, 3H),3.65 (t, 2H), 1.60 (s, 3H).

d)(S)-3-[4-(5-Difluoromethyl-6-methoxy-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

A solution of triphenylphosphine (74 mg, 0.28 mmol) in THF (2 ml) wastreated with DEAD (0.04 ml, 0.28 mmol) followed by(R)-3-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (CASregistry 127423-61-4) (45 mg, 0.24 mmol) and4-(5-difluoromethyl-6-methoxy-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-ol(65 mg, 0.20 mmol). The resulting red/brown solution was stirred at 70°C. for 18 h, cooled down to rt, diluted with EtOAc and washed with sat.aq. NaHCO₃ soln. The organic layer was dried over MgSO₄, concentratedand purified by flash chromatography on silica gel (cyclohexane/EtOAc100:0 to 70:30) to afford the title compound as a yellow solid (51 mg,42% yield).

HPLC Rt_(M1)=1.36 min; ESIMS: 492 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.92-7.75 (m, 1H), 7.46-7.33 (m, 1H),7.19-6.84 (t, 1H), 6.77 (s, 2H), 4.86 (br s, 1H), 3.98 (br s, 2H), 3.88(s, 3H), 3.73-3.55 (m, 2H), 3.46-3.33 (m, 2H), 2.12-1.95 (m, 2H),1.65-1.55 (m, 3H), 1.34 (br s, 9H). Rotamers.

e){(S)-3-[4-(5-Difluoromethyl-6-methoxy-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidin-1-yl}-(1,1-dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-methanone

A solution of(S)-3-[4-(5-difluoromethyl-6-methoxy-pyridin-3-yl)-5-methyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (51 mg, 0.10 mmol) in DCM (3 ml) was treated underargon at rt with TFA (0.08 ml, 1.10 mmol) and stirred for 18 h. Thereaction mixture was quenched with sat. aq. NaHCO₃ soln. and the organicsolution was separated through a phase separating affording a yellowsolution. 1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-carboxylic acid(CAS registry 64096-87-3) (25 mg, 0.14 mmol), Et₃N (0.05 ml, 0.33 mmol),EDC (31 mg, 0.16 mmol) and HOBT (25 mg, 0.16 mmol) were added and thereaction mixture was stirred at rt for 3 h. The reaction mixture wasquenched with sat. aq. NaHCO₃ soln. The organic layer was separated byelution through a phase separating cartridge, the crude product waspurified over SFC (column Reprosil NH₂ (250×30 mm (l×w), 60 A, 5 μm,Princeton, gradient of methanol in supercritical CO₂) to afford thetitle compound as a yellow oil (19 mg, 30% yield)

HPLC Rt_(M1)=1.00 min; ESIMS: 552 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆, 375K): δ 7.85 (br s, 1H), 7.43 (br s, 1H),6.96 (t, 1H), 6.80 (s, 2H), 5.11-4.71 (m, 1H), 4.04 (t, 2H), 3.94 (s,3H), 3.70 (d, 2H), 3.63 (br s, 2H), 3.52 (br s, 2H), 3.23-3.04 (m, 4H),2.92-2.73 (m, 1H), 2.16 (br s, 2H), 2.06 (br s, 4H), 1.67 (s, 3H),Rotamers.

Example Y2-Methoxy-5-{6-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-ylamino]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile

a) 5-Iodo-2-methoxy-nicotinonitrile

A mixture of 2-methoxy-nicotinonitrile (CAS registry 7254-34-4) (10 g,74.6 mmol) and N-iodosuccinimide (CAS registry 516-12-1) (25.2 g, 112mmol) was treated with trifluoroacetic acid (CAS registry 76-05-1) (68.9ml, 895 mmol) and trifluoroacetic anhydride (CAS registry 407-25-0)(31.6 ml, 224 mmol) and the reaction mixture was heated at 90° C. for 18h then cooled to rt and poured onto ice. The mixture was slowly basifiedusing 30% aq. NaOH soln., diluted with water and extracted with EtOAc.The organic layer was successively washed with 20% aq. sodiumthiosulfate soln., and sat. aq. NaHCO₃ soln., dried over Na₂SO₄,filtered and concentrated. The title compound was obtained after flashchromatography on silica gel (cyclohexane/EtOAc, 100:0 to 20:80) as asolid (12.2 g, 63% yield)

UPLC Rt_(M14)=1.30 min; ESIMS: 261 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.54 (d, 1H), 8.11 (d, 1H), 4.06 (s, 3H).

b)5-(6-Bromo-2,3-dihydro-benzo[1,4]oxazin-4-yl)-2-methoxy-nicotinonitrile

A mixture of 6-bromo-3,4-dihydro-2H-benzo[1,4]oxazine (CAS registry105655-01-4) (5.0 g, 23.36 mmol), 5-iodo-2-methoxy-nicotinonitrile (12.2g, 46.7 mmol) and NaOtBu (2.69 g, 28.0 mmol) in toluene (50 ml) wasdegassed with argon for 10 min, thenbis(tri-tert-butylphosphine)-palladium(0) (0.36 g, 0.70 mmol) was added.The reaction mixture stirred at 110° C. for 18 h under argon. Aftercooling to rt, the reaction mixture was filtered through celite, rinsedwith EtOAc and the filtrates were washed with sat. aq. NaHCO₃ soln. Theorganic layer was dried over Na₂SO₄, filtered, concentrated and purifiedby flash chromatography on silica gel (cyclohexane/EtOAc, 100:0 to50:50) to yield the title compound (4.2 g, 52% yield).

UPLC Rt_(M14)=1.55 min; ESIMS: 348 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.31 (d, 1H), 7.80 (d, 1H), 6.89 (dd, 1H),6.78 (d, 1H), 6.67 (d, 1H), 4.30-4.35 (m, 2H), 4.10 (s, 3H), 3.61-3.66(m, 2H).

c)(S)-3-[4-(5-Cyano-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester

A mixture of5-(6-bromo-2,3-dihydro-benzo[1,4]oxazin-4-yl)-2-methoxy-nicotinonitrile(300 mg, 0.87 mmol), (S)-3-amino-pyrrolidine-1-carboxylic acidtert-butyl ester (CAS registry 147081-44-5) (0.26 ml, 1.47 mmol),2-(dicyclohexylphosphino)biphenyl (CAS registry 247940-06-3) (18.2 mg,0.05 mmol) and NaOtBu (100 mg, 1.04 mmol) in toluene (10 ml) wasdegassed with argon for 10 min, then Pd₂(dba)₃ (23.8 mg, 0.03 mmol) wasadded. The reaction mixture was stirred at 110° C. for 1 h under argon.After cooling to rt, the reaction mixture was filtered through celite,rinsed with EtOAc and the filtrates were concentrated. The titlecompound was obtained after flash chromatography on silica gel(cyclohexane/EtOAc, 100:0 to 70:30) as a yellow foam (150 mg, 37%yield).

UPLC Rt_(M11)=2.73 min; ESIMS: 452 [(M+H)⁺].

¹H NMR (400 MHz, CD₃OD): δ 8.36 (d, 1H), 8.04 (d, 1H), 6.68 (s, 1H),6.19 (dd, 1H), 6.03 (s, 1H), 4.19-4.24 (m, 2H), 4.06 (s, 3H), 3.81-3.89(m, 1H), 3.62-3.68 (m, 2H), 3.48-3.56 (m, 1H), 3.35-3.48 (m, 2H),3.08-3.18 (m, 1H), 2.05-2.18 (m, 1H), 1.75-1.87 (m, 1H), 1.46 (s, 9H).

d)2-Methoxy-5-[6-((S)-pyrrolidin-3-ylamino)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-nicotinonitrile

A solution of(S)-3-[4-(5-cyano-6-methoxy-pyridin-3-yl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester (144 mg, 0.32 mmol) in CH₂Cl₂ (4 ml) was treatedwith TFA (0.49 ml, 6.38 mmol). The reaction mixture was stirred at rtfor 2 h, quenched with sat. aq. NaHCO₃ soln. and extracted with DCM. Theorganic layer was dried over Na₂SO₄, filtered and concentrated underreduced pressure to afford the title product as a yellow foam (120 mg,100% yield).

UPLC Rt_(M11)=2.06 min; ESIMS: 352 [(M+H)⁺].

¹H NMR (400 MHz, CD₃OD): δ 8.36 (d, 1H), 8.04 (d, 1H), 6.69 (d, 1H),6.17 (dd, 1H), 6.00 (d, 1H), 4.19-4.26 (m, 2H), 4.06 (s, 3H), 3.80-3.90(m, 1H), 3.62-3.69 (m, 2H), 3.10-3.20 (m, 2H), 2.98-3.09 (m, 1H),2.82-2.90 (m, 1H), 2.06-2.18 (m, 1H), 1.70-1.81 (m, 1H).

e)2-Methoxy-5-{6-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-ylamino]-2,3-dihydro-benzo[1,4]oxazin-4-yl}-nicotinonitrile

At 0° C., a solution of2-methoxy-5-[6-((S)-pyrrolidin-3-ylamino)-2,3-dihydro-benzo[1,4]oxazin-4-yl]-nicotinonitrile(27 mg, 0.08 mmol) in CH₂Cl₂ (1 ml) was treated with Et₃N (0.02 ml, 0.12mmol) and tetrahydro-pyran-4-carbonyl chloride (CAS registry 40191-32-0)(11 μl, 0.09 mmol). The reaction mixture was stirred at 0° C. for 1 h,then quenched with sat. aq. NaHCO₃ soln. and extracted with DCM. Theorganic layer was dried over Na₂SO₄, filtered and concentrated and thetitle compound was obtained after prep. RP-HPLC (column Sunfire PrepC18OBD 30×100 mm, 5 μm; solvent A: H₂O+0.1 Vol.-% TFA; solvent B: CH₃CN+0.1Vol.-% TFA, gradient 5-60% B in 20 min) and filtration over AgilentPL-HCO₃ MP SPE cartridge as a yellow solid (9 mg, 25% yield).

UPLC Rt_(M2)=1.19 min; ESIMS: 464 [(M+H)⁺].

¹H NMR (400 MHz, CD₃OD): δ 8.36 (d, 1H), 8.05 (m, 1H), 6.69 (dd, 1H),6.15-6.23 (m, 1H), 5.97-6.07 (m, 1H), 4.15-4.29 (m, 2H), 4.06 (s, 3H),3.82-4.03 (m, 3H), 3.35-3.80 (m, 8H), 2.65-2.84 (m, 1H), 2.03-2.28 (m,1H), 1.49-2.03 (m, 5H).

Coupling Conditions

A) Buchwald Aminations or Hydroxylations

Solvents Typical Typical Condition # Pd source Ligand Base usedtemperature reaction time CA1 Pd₂(dba)₃ Rac-BINAP Cs₂CO₃ toluene 60-100°C. 16 h CA2 Pd₂(dba)₃ XPhos NaO^(t)Bu dioxane 100° C. 1-18 h CA3Pd₂(dba)₃ XPhos Cs₂CO₃ toluene 100° C. CA4 Pd₂(dba)₃ XPhos Cs₂CO₃dioxane 100° C. 1-18 h CA5 Pd₂(dba)₃ tetramethyl- KOH dioxane/ 100° C.18-72 h t-butyl- H₂O XPhos CA6 Pd₂(dba)₃ XPhos NaO^(t)Bu toluene 80-110°C. 20 min-18 h CA7 Pd(P(tBu₃)₂ NaO^(t)Bu toluene 140° C. (mw) 30 min CA8Pd(P(tBu₃)₂ NaO^(t)Bu toluene 120° C. 18 h CA9 Pd₂(dba)₃ XPhos NaO^(t)Butoluene 125° C. (mw) 25 min CA10 Pd(OAc)₂ Rac-BINAP Cs₂CO₃ toluene60-100° C. 21 h CA11 Pd[Ruphos] RuPhos NaO^(t)Bu dioxane 100° C. 18 hCA12 Pd(P(tBu₃)₂ K₃PO₄ toluene 120° C. 18 h CA13 Pd₂(dba)₃ XPhos K₃PO₄toluene 100° C. 12 h CA14 Pd₂(dba)₃ XPhos K₃PO₄ toluene 110-120° C.  12h CA15 Pd(P(tBu₃)₂ NaO^(t)Bu THF 110° C. 18 h CA16 Pd₂(dba)₃ XPhosNaO^(t)Bu THF 110° C.   1 h-18 h

B) Amide Bond Formation Conditions

Condi- Coupling Solvents Typical Typical tion# reagents used temperaturereaction time CB1 HBTU DMF or DMA rt 1 h-18 h CB2 HOBT, EDC DMF rt 1h-18 h CB3 HATU CH₂Cl₂ 0° C. to rt 30 min CB4 HOBT, EDC CH₂Cl₂ rt 18 h CB5 COMU, DIPEA DMF rt 12 h  CB6 none CH₂Cl₂ rt 1 h-18 h CB7 HBTU CH₂Cl₂rt 30 min

C) Side Chain Introduction Conditions

CC1) Using Mesylate

At rt, a dry solution of pyridin-2-ol intermediate (1 eq.) and mesylateintermediate (1.1-2 eq.) in DMF (0.17 M) was treated with NaH in mineraloil (2-3 eq.) and the reaction mixture was stirred at 20-80° C. for 4 to72 h.

CC2) Using Mesylate

At rt, a dry solution aryl-6-ol intermediate (1 eq.) and mesylateintermediate (1.1-2 eq.) in DMF (0.17M) was treated with NaH in mineraloil (2-3 eq.) and the reaction mixture was stirred at 50-80° C. for 4 to72 h.

CC3) Using Mesylate

At rt, a dry solution of aryl-6-ol intermediate (1 eq.) and mesylateintermediate (2.5 eq.) in DMF (0.06M) was treated with K₂CO₃ (4 eq.) andthe reaction mixture was stirred at 85° C. to 100° C. for 4 to 50 h.

CC4) Using Mitsunobu

At rt, DEAD (1.4 eq.), (R)-3-Hydroxy-pyrrolidine-1-carboxylic acidtert-butyl ester (1.5 eq.) and aryl-6-ol intermediate (1 eq.) were addedto a solution of triphenylphosphine (1.4 eq.) in THF (0.30M). Thered/brown solution was stirred at 70° C. for 18 h.

D) Conditions for Chiral Separation Chromatography

Meth- UV De- od# Column Eluent tection CD1 Chiralpak ICn-Heptane/DMME/IPA/DEA 230 nm 250 × 30 mm, 5 μm 20:50:30:0.05 CD2Chiralpak IC ACN 100% 230 nm 250 × 30 mm, 5 μm CD3 Chiralcel ODHEtOH/MeOH 60:40 220 nm 250 × 30 mm, 5 μm CD4 Chiralpak ICDMME/IPA/MeOH/DEA 230 nm 250 × 30 mm, 5 μm 70:25:5:0.05, CD5 ChiralpakIC ACN 100% 220 nm 250 × 30 mm, 5 μm CD6 Chiralpak IC MeOH 100% 220 nm250 × 30 mm, 5 μm CD7 Chiralpak IC n-Heptane/DMME/EtOH/DEA 240 nm 250 ×46 mm, 5 μm 40:50:10:0.05 CD8 Chiralcel OD-H, CO₂/IPA 70:30 (isocratic215 nm 4.6 × 250 mm CD9 Chiralpak AD-H, Heptane/EtOH 60:40 5 μm CD10Chiralpak IC DMME/IPA/MeOH/DEA 230 nm 250 × 46 mm, 5 μm 70:25:5:0.05,CD11 Chiralcel ODH EtOH/MeOH 60:40 220 nm 250 × 20 mm, 5 μm CD12Chiralpak IC EtOH/MeOH 50:50 210 nm 765 × 37.5 cm, 20 μm

Preparation of Intermediates

IA) Aromatic Bromides

IA) Aromatic Bromides Comment on Intermediate # Structure Autonom namesynthesis IA1

5-Bromo-2- methanesulfonyl-3-methyl- pyridine 1 step from CAS1289270-74-1 IA2

5-Bromo-3-fluoro-2- methanesulfonyl- pyridine 1 step from CAS1289007-85-7 IA3

5-Bromo-2- methanesulfonyl-3- trifluoromethyl-pyridine 1 step from CAS211122-42-8 IA4

5-Bromo-3-difluoromethyl- 2-methanesulfonyl- pyridine 2 steps from CAS852181-11-4 IA5

5-Bromo-3-fluoromethyl-2- methanesulfonyl-pyridine 2 steps from CAS742100-75-0 IA6

5-Bromo-3-difluoromethyl- 2-methoxy-pyridine CAS 1254123-51-7 IA7

5-Bromo-3-fluoromethyl-2- methoxy-pyridine 1 step from CAS 351410-47-4IA8

5-Bromo-2- difluoromethoxy-3-methyl- pyridine CAS 1214337-94-6 IA9

5-Bromo-2-methoxy-3- methyl-pyridine CAS 760207-87-2 IA10

5-Bromo-3-fluoro-2- methoxy-pyridine CAS 124432-70-8 IA11

5-Bromo-3-chloro-2- methoxy-pyridine CAS 848366-28-9 IA12

5-Bromo-2-methoxy- nicotinonitrile CAS 941294-54-8 IA13

5-Bromo-pyridine-3- sulfonic acid dimethylamide CAS 896160-99-9 IA14

4-(5-Bromo-pyridine-3- sulfonyl)-morpholine CAS 889676-35-1 IA15

5-Bromo-2-methyl-3-nitro- pyridine CAS 911434-05-4 IA16

5-Bromo-2-methyl-pyridine CAS 3430-13-5 IA17

5-Bromo-nicotinonitrile CAS 35590-37-5 IA18

5-bromo- trifluoromethylpyridine CAS 436799-33-6 IA19

5-Bromo-2-ethoxy-3- methyl-pyridine CAS 610279-03-3 IA20

5-bromo-2- methoxypyridine CAS 13472-85-0 IA21

5-bromo-2-methoxy-3- trifluoromethylpyridine CAS 1214377-42-0 IA22

5-Bromo-2-methoxy- nicotinic acid methyl ester CAS 122433-41-4 IA23

2-amino-5-bromo-3- trifluoromethylpyridine CAS 79456-34-1 IA24

1-(5-Bromo-2-methoxy- pyridine-3-sulfonyl)-4- methyl-piperazine IA25

5-Bromo-2- methoxymethyl-pyridine CAS 1000787-43-8 IA26

5-bromo-2- trifluromethylpyridine CAS 436799-32-5 IA27

5-Bromo-3-methyl-pyridin- 2-ylamine CAS 3430-21-5 IA28

5-Bromo-pyridine-2- carbonitrile CAS 97483-77-7 IA29

5-Bromo-2,3-dimethoxy- pyridine CAS 52605-98-8 IA30

3-Bromo-5-(propane-2- sulfonyl)-pyridine IA31

1-(5-Bromo-2-methyl- benzenesulfonyl)- piperidine CAS 364736-61-8 IA32

4-(5-Bromo-2-methoxy- benzenesulfonyl)- morpholine CAS 325809-68-5 IA33

4-(5-Bromo-pyridine-3- sulfonyl)-morpholine CAS 889676-35-1 IA34

1-(5-Bromo-2-methoxy- benzenesulfonyl)-4- methyl-piperazine CAS325809-71-0 IA35

1-(5-Bromo-pyridine-3- sulfonyl)-4-methyl- piperazine CAS 1007212-08-9IA36

5-Bromo-2,N-dimethoxy- N-methyl- benzenesulfonamide CAS 1247891-51-5IA37

5-Bromo-pyridine-3- sulfonic acid methoxy- methyl-amide CAS 1248282-54-3IA38

3-Bromo-5-methoxy- pyridine CAS 50720-12-2 IA39

3-Bromo-5-chloro-pyridine CAS 73583-39-8 IA40

5-Bromo-2- methanesulfonyl-pyridine CAS 98626-95-0 IA41

5-Bromo-pyridine-3- sulfonic acid dimethylamide CAS 896160-99-9 IA42

5-Bromo-pyridin-3-ylamine CAS 13535-01-8 IA43

5-Bromo-pyridine-3- sulfonic acid ethylamide CAS 1065074-78-3 IA44

5-Bromo-2-ethanesulfinyl- pyridine IA45

5-Bromo-2- methanesulfonyl-3- methoxy-pyridine IA46

5-Bromo-3-methoxy- pyridin-2-ylamine CAS 42409-58-5 IA47

N-(5-Bromo-2-methoxy- pyridin-3-yl)- methanesulfonamide CAS 1083327-58-5IA48

5-Bromo-3-ethyl-2- methoxy-pyridine CAS 1256788-92-7 IA49

5-Bromo-2-chloro-3- methoxy-pyridine CAS 286947-03-3 IA50

5-Bromo-2-chloro-3- methyl-pyridine CAS 29241-60-9 IA51

(5-Bromo-2- methanesulfonyl-pyridin-3- yl)-methyl-amine IA52

(5-Bromo-2- methanesulfonyl-pyridin-3- yl)-dimethyl-amine IA53

5-Bromo-3-chloro-2- methanesulfonyl- pyridine 1335052-54-4 IA54

3-Bromo-quinoline CAS 5332-24-1 IA55

3-Bromo-5- methanesulfonyl-pyridine CAS 445491-71-4 IA56

5-Bromo-2-methyl- benzonitrile CAS 156001-51-3 IA57

4-Bromo-1-methoxy-2- trifluoromethyl-benzene CAS 1514-11-0 IA58

5-Bromo-2-ethanesulfonyl- pyridine CAS 223556-06-7 IA59

4-Bromo-2- methanesulfonyl-1- methoxy-benzene CAS 90531-99-0 IA60

5-Bromo-2-methoxy-N,N- dimethyl- benzenesulfonamide CAS 871269-16-8 IA61

4-Bromo-2-methyl-pyridine CAS 22282-99-1 IA62

4-Bromo-2-methoxy- pyridine CAS 100367-39-3 IA63

4-Bromo-2-trifluoromethyl- pyridine CAS 887583-90-6 IA64

4-Bromo-pyridine-2- carbonitrile CAS 62150-45-2 IA65

5-Bromo-2,N-dimethoxy- benzenesulfonamide IA66

4-Bromo-1,2-dimethoxy- benzene CAS 2859-78-1 IA67

5-Bromo-2-methyl-pyridin- 3-ylamine CAS 914358-73-9 IA68

(5-Bromo-2-methyl- pyridin-3-yl)-dimethyl- amine CAS 1280592-37-1 IA69

2-(Benzyloxy)-5-bromo-3- methylpyridine CAS 1289270-73-0 IA70

5-Bromo-3- (dimethoxymethyl)-2- methoxypyridine CAS 351410-59-8

Intermediate IA1 5-bromo-2-methanesulfonyl-3-methyl-pyridine

A solution of 5-bromo-2-methylsulfanyl-3-methyl-pyridine (9.04 g, 41.4mmol) in DCM (83 ml) was treated at 0° C. with mCPBA (21.46 g, 124mmol). After stirring for 18 h at rt, the reaction mixture was added to2N aq. NaOH soln. and was extracted with DCM. The organic layer wasdried over Na₂SO₄, concentrated and the title compound was obtainedafter trituration with cyclohexane to afford a white solid (9.25 g, 89%yield).

UPLC Rt_(M1)=0.81 min; MS (ESI, m/z): 250.1 [(M+H⁺]

¹H NMR (400 MHz, DMSO-d₆): δ 8.68 (d, 1H), 8.30 (d, 1H), 3.37 (s, 3H),2.58 (s, 3H).

Intermediate IA2 5-bromo-3-fluoro2-methanesulfonyl-pyridine

A solution of 5-bromo-3-fluoro-2-methylsulfanyl-pyridine (222 mg, 1.0mmol) in DCM (5 ml) was treated at 0° C. with mCPBA (518 mg, 3.0 mmol).After stirring for 1.5 h at rt the reaction mixture was added to 2N aq.NaOH soln. and was extracted with DCM. The organic layer was dried overNa₂SO₄, concentrated and the title compound was obtained as a whitesolid (241 mg, 95% yield) which was used without further purification.

UPLC Rt_(M1)=0.63 min;

¹H NMR (400 MHz, DMSO-d₆): δ 8.75 (d, 1H), 8.60 (d, 1H), 3.40 (s, 3H).

Intermediate IA3 5-Bromo-2-methanesulfonyl-3-trifluoromethyl-pyridine

A solution of 5-bromo-2-methylsulfanyl-3-trifluoromethyl-pyridine (1.40g, 1.16 mmol) in DCM (30 ml) was treated at 0° C. with mCPBA (2.67 g,15.48 mmol). After stirring for 18 h at rt the reaction mixture wasadded to a 4N aq. NaOH soln. and was extracted with DCM. The organiclayer was dried over Na₂SO₄, filtered, concentrated and the titlecompound was obtained after flash chromatography on silica gel(hexane/EtOAc, 100:0 to 70:30) as a white solid (940 mg, 60% yield).

UPLC Rt_(M1)=0.87 min; MS (ESI, m/z): 323.0 [(M+NH₄)⁺].

¹H NMR (400 MHz, CDCl₃): δ 9.45 (d, 1H), 8.76 (d, 1H), 3.57 (s, 3H).

Intermediate IA4 5-Bromo-3-difluoromethyl-2-methanesulfonyl-pyridine a)2,5-Dibromo-3-difluoromethyl-pyridine

A solution of Et₃N trifluoride (1.80 ml, 11.3 mmol) in DCM (40 ml) wastreated at 0° C. with Xtalfluor-E (2.67 g, 15.5 mmol) and2,5-dibromo-pyridine-3-carbaldehyde (1.0 g, 3.77 mmol). After stirringfor 19 h at rt the reaction mixture was diluted with TBME and washedwith sat. aq. NaHCO₃ soln. The organic layer was dried over Na₂SO₄,concentrated to leave a yellow oil (950 mg, 88% yield) which was usedwithout further purification.

UPLC Rt_(M1)=1.05 min;

¹H NMR (400 MHz, DMSO-d₆): δ 8.57 (d, 1H), 8.48 (d, 1H), 7.14 (t, 1H),3.35 (s, 3H).

b) 5-Bromo-3-difluoromethyl-2-methanesulfonyl-pyridine

A solution of 2,5-dibromo-3-difluoromethyl-pyridine (1400 mg, 1.16 mmol)in DMF (10 ml) was treated at 0° C. with sodium methanethiolate (348 mg,4.97 mmol). After stirring for 1.5 h at rt, the reaction was cooled at0° C. and mCPBA (2857 mg, 16.56 mmol) was added to the reaction mixture.After stirring for 1 h at rt the reaction mixture was added to a 4N aq.NaOH soln. and was extracted with TBME. The organic layer was dried overNa₂SO₄, filtered, concentrated and the title compound was obtained afterflash chromatography on silica gel (hexane/EtOAc, 100/0 to 80/20) as awhite solid (498 mg, 53% yield).

UPLC Rt_(M1)=0.86 min;

¹H NMR (400 MHz, CDCl₃): δ 8.67 (d, 1H), 8.45 (d, 1H), 7.37 (t, 1H),3.37 (s, 3H).

Intermediate IA5 5-Bromo-3-fluoromethyl-2-methanesulfonyl-pyridine a)5-Bromo-2-chloro-3-fluoromethyl-pyridine

A solution of Et₃N trifluoride (1.76 ml, 10.79 mmol) in DCM (20 ml) wastreated at 0° C. with Xtalfluor-E (1.65 g, 7.19 mmol) and(5-bromo-2-chloro-pyridin-3-yl)-methanol (0.80 g, 3.60 mmol). Afterstirring for 18 h at rt the reaction mixture was diluted with TBME andwashed with sat. aq. NaHCO₃ soln. The organic layer was dried overNa₂SO₄, concentrated and the title compound was obtained after flashchromatography on silica gel (hexane/EtOAc, 100/0 to 90/10) as acolourless oil (286 mg, 35% yield).

UPLC Rt_(M1)=0.98 min;

¹H NMR (400 MHz, CDCl₃): δ 8.53 (d, 1H), 8.45 (d, 1H), 5.52 (d, 1H)

b) 5-Bromo-3-fluoromethyl-2-methanesulfonyl-pyridine

A solution of 2,5-dibromo-3-difluoromethyl-pyridine (1.4 g, 1.16 mmol)in DMF (10 ml) was treated at 0° C. with sodium methanethiolate (348 mg,4.97 mmol). After stirring at rt for 1.5 h, the reaction was cooled downto 0° C. and mCPBA (2857 mg, 16.56 mmol) was introduced to the reactionmixture. After stirring for 1 h at rt the reaction mixture was added toan 4N aq. NaOH soln. and was extracted with TBME. The organic layer wasdried over Na₂SO₄, filtered, concentrated and the title compound wasobtained after flash chromatography on silica gel (hexane/EtOAc, 100/0to 80/20) as a white solid (498 mg, 53% yield) UPLC Rt_(M1)=0.86 min;

¹H NMR (400 MHz, CDCl₃): δ 8.67 (d, 1H), 8.45 (d, 1H), 7.37 (t, 1H),3.37 (s, 3H).

Intermediate IA7 5-Bromo-3-fluoromethyl-2-methoxy-pyridine

A solution of (5-bromo-2-methoxy-pyridin-3-yl)-methanol (343 mg, 1.57mmol) in DCM (7 ml) was treated at 0° C. with deoxofluor (1.5 ml, 3.46mmol) and EtOH (27.6 μl, 0.47 mmol). The reaction mixture was stirred atrt, quenched with sat. aq. NaHCO₃ soln. and was extracted with DCM. Theorganic layer was dried over Na₂SO₄, concentrated to afford the titlecompound after flash chromatography on silica gel (hexane/EtOAc 80:20)as a yellow oil (80 mg, 23% yield).

UPLC Rt_(M1)=1.07 min;

¹H NMR (400 MHz, DMSO-d₆): δ 8.32 (t, 1H), 8.00 (t, 1H), 5.37 (d, 2H),3.89 (s, 3H).

Intermediate IA241-(5-Bromo-2-methoxy-pyridine-3-sulfonyl)-4-methyl-piperazinea)(5-Bromo-2-methanesulfonyl-pyridin-3-yl)-methyl-amine

A solution of 5-bromo-2-chloro-pyridine-3-sulfonyl chloride (CASregistry 1146290-19-8) (580 mg, 1.99 mmol) and Et₃N (0.55 ml, 3.99 mmol)in DCM (20 ml) at 0° C. was treated with 1-methyl-piperazine (0.33 ml,2.99 mmol), the resulting mixture was stirred at 0° C. for 20 min and atrt for 18 h. The mixture was diluted with DCM (30 ml) and washed withsat.aq. NaHCO₃ soln. The organic layer was dried over MgSO₄,concentrated and purified by flash chromatography on silica gel(DCM/MeOH 100:0 to 90:10) to afford the title compound as an orangesolid (420 mg, 59% yield).

HPLC Rt_(M2)=1.49 min; ESIMS: 356 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.62 (d, 1H), 8.50 (d, 1H), 3.57-3.29 (t,4H), 2.50 (t, 4H), 2.33 (s, 3H).

b) 1-(5-Bromo-2-methoxy-pyridine-3-sulfonyl)-4-methyl-piperazine

A solution of1-(5-bromo-2-chloro-pyridine-3-sulfonyl)-4-methyl-piperazine (420 mg,1.18 mmol) in THF (10 ml) was treated portionwise with sodium methoxide(192 mg, 3.55 mmol). The resulting mixture was stirred at rt for 18 h.The mixture was quenched by addition of water and extracted with EtOAc.The combined organic layers were washed with sat. aq. NaHCO₃ soln. anddried over MgSO₄, concentrated and purified by flash chromatography onsilica gel (DCM/MeOH 100:0 to 90:10) to afford the title compound (358mg, 85% yield).

HPLC Rt_(M2)=1.47 min; ESIMS: 350, 352 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.36 (d, 1H), 8.28 (d, 1H), 4.05 (s, 3H),3.35 (br s, 4H), 2.49 (br s, 4H), 2.33 (s, 3H).

Intermediate IA30 3-Bromo-5-(propane-2-sulfonyl)-pyridine

A solution of 3,5-dibromo-pyridine (CAS registry 625-92-3) (495 mg, 2.09mmol) in NMP (5 ml) was treated with sodium propane-2-thiolate (CASregistry 20607-43-6) (205 mg, 2.09 mmol), the resulting mixture wasstirred at 80° C. for 2 h. The mixture was cooled down and diluted withEtOAc, washed with water (2×), then brine. The organic layer was driedover MgSO₄ and concentrated. The obtained residue was dissolved in DCM(10 ml), treated with mCPBA (1083 mg, 6.27 mmol) and stirred at rt for18 h. 10% aq. sodium sulfite soln. was added and the mixture was stirredat rt for 1 h. The phases were separated and the organic layer waswashed with sat. aq. NaHCO₃ soln., dried over MgSO₄, concentrated andpurified by flash chromatography on silica gel (cyclohexane/EtOAc 100:0to 00:100) to afford the title compound (364 mg, 59% yield).

HPLC Rt_(M2)=0.77 min; ESIMS: 264, 266 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 8.98 (dd, 2H), 8.33 (t, 1H), 3.34-3.17 (m,1H), 1.37 (d, 6H).

Intermediate IA44 5-Bromo-2-ethanesulfinyl-pyridine a)5-Bromo-2-ethylsulfanyl-pyridine

A mixture of 2,5-dibromo-pyridine (CAS registry 588729-99-1) (1.15 g,4.85 mmol) and sodium ethanthiolate (CAS registry 811-51-8) (2.04 g,24.27 mmol) in DMSO (15 ml) was stirred at rt for 18 h. Water was addedand the mixture was extracted with DCM. The organic layer was dried bypassing it through a phase separating cartridge, was concentrated andpurified by flash chromatography on silica gel (cyclohexane/EtOAc 100:0to 70:30) to afford the title compound as an orange oil (1.09 g, 92%yield).

HPLC Rt_(M2)=1.25 min; ESIMS: 218, 220 [(M+H)⁺].

b) 5-Bromo-2-ethanesulfinyl-pyridine

A solution of 5-Bromo-2-ethylsulfanyl-pyridine (1.09 g, 4.49 mmol) inDCM (15 ml) was treated with mCPBA (1.55 g, 8.98 mmol). The resultingsolution was stirred at rt for 15 min. The mixture was quenched byaddition of aq. 2M NaOH soln., and extracted with DCM. The combinedorganic layers were dried over MgSO₄ and concentrated. The compound waspurified by prep. RP-HPLC (column SunFire C18 OBD, gradient 5-60% ACN in15 min) to afford the title compound as a colourless oil (490 mg, 45%yield).

HPLC Rt_(M2)=0.65 min; ESIMS: 234, 236 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.85 (d, 1H), 8.37 (dd, 1H), 7.80 (d, 1H),3.33 (s, 2H), 3.27-3.10 (m, 1H), 2.94-2.72 (m, 1H), 1.01 (t, 3H).

Intermediate IA45 5-Bromo-2-methanesulfonyl-3-methoxypyridine a)5-Bromo-3-methoxy-2-methylsulfanyl-pyridine

A mixture of 2,5-dibromo-3-methoxy-pyridine (CAS registry 1142191-57-8)(550 mg, 2.06 mmol) and sodium methanethiolate (CAS registry5188-07-8)(722 mg, 10.30 mmol) in DMSO (1.5 ml) was stirred at rt for0.5 h. Water was added and the mixture was extracted with DCM. Theorganic layer was dried by passing it through a phase separatingcartridge and was concentrated to afford the title compound as acolourless oil (1.50 g, 93% yield, crude).

¹H NMR (400 MHz, DMSO-d₆): δ 8.19 (d, 1H), 7.53 (d, 1H), 3.89 (s, 3H),3.32 (s, 3H).

b) 5-Bromo-2-methanesulfonyl-3-methoxypyridine

A solution of 5-bromo-3-methoxy-2-methylsulfanyl-pyridine (1.50 g, 2.24mmol) in DCM (10 ml) was treated with mCPBA (1.55 g, 8.97 mmol). Theresulting mixture was stirred at rt for 18 h. The mixture was quenchedby addition of aq. 2M NaOH soln., and extracted with DCM. The organiclayer was dried by passing through a phase separating cartridge andconcentrated to afford the title compound (400 mg, 33% yield as crude).

HPLC Rt_(M2)=0.75 min; ESIMS: 268 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.41 (d, 1H), 8.17 (s, 1H), 4.01 (s, 3H),3.30 (s, 3H).

Intermediate IA51 (5-Bromo-2-methanesulfonyl-pyridin-3-yl)-methyl-aminea) (5-Bromo-2-chloro-pyridin-3-yl)-methyl-amine

A solution of 5-bromo-2-chloro-pyridin-3-ylamine (CAS registry588729-99-1) (565 mg, 2.72 mmol) in THF (4 ml) at 0° C. was treated withBuLi 1.6M in hexane (0.17 ml, 0.17 mmol), the resulting mixture wasstirred at 0° C. for 0.5 h, then methyl iodide (0.17 ml, 2.72 mmol) wasslowly added. The reaction mixture was allowed to warm to rt and wasstirred for 18 h. The orange/brown mixture was poured into sat. aq.NaHCO₃ soln., and extracted with EtOAc. The organic layer was dried overMgSO₄, concentrated and purified by flash chromatography on silica gel(cyclohexane/EtOAc 95:5 to 60:40) to afford the title compound as anorange solid (354 mg, 59% yield).

HPLC Rt_(M1)=0.94 min; ESIMS: 221, 223, 225 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.65 (d, 1H), 7.14 (d, 1H), 6.11 (d, 1H),2.74 (d, 3H).

b) (5-Bromo-2-methanesulfonyl-pyridin-3-yl)-methyl-amine

A solution of (5-bromo-2-chloro-pyridin-3-yl)-methyl-amine (354 mg, 1.60mmol) in DMF (2.6 ml) was treated with sodium methanethiolate (168 mg,2.40 mmol) at 0° C. The resulting solution was stirred at rt for 2 d andheated at 60° C. for 4 d. The mixture was quenched at 0° C. by additionof aq. 2M NaOH soln., and extracted with DCM. The combined organiclayers were dried over MgSO₄ and concentrated to afford an orange oilwhich was dissolved in DCM (5 ml) and treated at 0° C. with mCPBA (CASregistry 937-14-4) (827 mg, 4.79 mmol) and stirred for 18 h at rt. Themixture was quenched at 0° C. by addition of aq. NaOH 2M soln. andextracted with DCM. Combined organics were dried over MgSO₄,concentrated and purified by flash chromatography on silica gel(cyclohexane/EtOAc 92:8 to 32:68) to afford the title compound as anorange solid (174 mg, 41% yield).

HPLC Rt_(M1)=0.78 min; ESIMS: 265, 267 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.95 (d, 1H), 7.50 (d, 1H), 6.70-6.59 (m,1H), 3.26 (s, 3H), 2.82 (d, 3H).

Intermediate IA52(5-Bromo-2-methanesulfonyl-pyridin-3-yl)-dimethyl-amine a)(5-Bromo-2-chloro-pyridin-3-yl)-methyl-amine

A solution of 5-bromo-2-chloro-pyridin-3-ylamine (CAS registry588729-99-1) (1 g, 4.82 mmol) in THF (7 ml) at 0° C. was treated withBuLi 1.6M in hexane (6 ml, 9.64 mmol), the resulting mixture was stirredat 0° C. for 0.5 h, then methyl iodide (0.60 ml, 9.64 mmol) was slowlyadded. The reaction mixture was allowed to warm at rt and was stirredfor 18 h. The orange/brown mixture was poured onto sat. aq. NaHCO₃soln., and extracted with EtOAc, the organic layer was dried over MgSO₄,concentrated and purified by flash chromatography on silica gel(cyclohexane/EtOAc 95:05 to 70:30) to afford the title compound as anorange solid (182 mg, 17% yield).

HPLC Rt_(M1)=0.94 min; ESIMS: 221, 223 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 7.65 (d, 1H), 7.14 (d, 1H), 6.11 (d, 1H),2.74 (d, 3H).

b) (5-Bromo-2-chloro-pyridin-3-yl)-dimethyl-amine

A solution of (5-bromo-2-chloro-pyridin-3-yl)-methyl-amine (182 mg, 0.82mmol) in DMF (4 ml) was treated with NaH (23 mg, 0.99 mmol) at rt, andthe mixture was stirred at rt for 0.5 h, methyl iodide (0.06 ml, 0.99mmol) was added and the resulting mixture was stirred at rt for 1 h. Themixture was diluted with TBME and washed with a sat. aq. NaHCO₃ soln.,the organic layer was dried over MgSO₄ and concentrated to afford thetitle compound as an orange oil (174 mg, 90% yield). It was used withoutfurther purification.

HPLC Rt_(M1)=1.03 min; ESIMS: 235, 237 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.10 (d, 1H), 7.70 (d, 1H), 2.78 (s, 6H).

c) (5-Bromo-2-methanesulfonyl-pyridin-3-yl)-methyl-amine

A solution of (5-bromo-2-chloro-pyridin-3-yl)-dimethyl-amine (174 mg,0.74 mmol) in DMF (5 ml) was treated with sodium methanethiolate (104mg, 1.48 mmol) at rt, the resulting solution was stirred at 80° C. for18 h. At 0° C., the mixture was quenched by addition of aq. 2M NaOHsoln. then extracted with TBME. The combined organic layers were driedover MgSO₄ and concentrated to afford an orange oil which was dissolvedin DCM (5 ml), treated at 0° C. with mCPBA (CAS registry 937-14-4) (382mg, 2.21 mmol) and stirred for 18 h at rt. At 0° C., the mixture wasquenched by addition of aq. 2M NaOH soln., then extracted with DCM. Thecombined organic layers were dried over MgSO₄, concentrated and purifiedby flash chromatography on silica gel (cyclohexane/EtOAc 88:12 to00:100) to afford the title compound as an orange solid (50 mg, 24%yield).

HPLC Rt_(M1)=0.83 min; ESIMS: 279, 281 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 8.25 (d, 1H), 7.90 (d, 1H), 3.29 (s, 3H),2.91 (s, 6H).

Intermediate IA65 5-Bromo-2,N-dimethoxy-benzenesulfonamide

A solution of 5-bromo-2-methoxy-benzenesulfonyl chloride (CAS registry23095-05-8) (218 mg, 0.76 mmol) and Et₃N (0.55 ml, 3.99 mmol) in DCM (20ml) at 0° C. was stirred for 15 min and treated withO-methylhydroxylamine (36 mg, 0.76 mmol), the resulting mixture wasstirred at rt for 18 h. The mixture was partitioned between EtOAc andwater. The organic layer was washed with sat.aq. NaHCO₃ soln., driedover MgSO₄ concentrated and purified by flash chromatography on silicagel (cyclohexane/EtOAc 100:0 to 0:100) to afford the title compound (189mg, 84% yield).

¹H NMR (400 MHz, CDCl₃): δ 8.10 (d, 1H), 7.78 (s, 1H), 7.72 (dd, 1H),6.97 (d, 1H), 4.01 (s, 3H), 3.81 (s, 3H).

IB) Carboxylic Acids or Acid Chlorides

IB) Carboxylic Inter- acids or acid mediate chlorides # StructureAutonom name IB1

Tetrahydro-furan-3- carboxylic acid (pure enantiomer) IB2

Tetrahydro-furan-3- carboxylic acid (pure enantiomer) IB3

5-tert- butoxycarbonylamino- 1-methyl-1H- imidazole-4-carboxylic acid

Example IB1/IB2 (S)-Tetrahydro-furan-3-carboxylicacid/(R)-Tetrahydro-furan-3-carboxylic acid a)Tetrahydro-furan-3-carboxylic acid benzyl ester

A solution of tetrahydro-furan-3-carboxylic acid (CAS registry89364-31-8) (4.00 g, 34.40 mmol) in DMF (20 ml) was treated with K₂CO₃(9.52 g, 68.9 mmol) and benzyl bromide (CAS registry 100-39-0) (8.18 ml,68.9 mmol) at 100° C. for 18 h. The mixture was cooled down to rt,diluted with EtOAc, washed with water and brine. Combined organic layerswere dried over MgSO₄, concentrated and purified by flash chromatographyon silica gel (cyclohexane/EtOAc 92:8 to 34:66) to afford the titlecompound as colourless oil (6.93 g, 98% yield)

HPLC Rt_(M2)=0.94 min; ESIMS: 207 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 7.50-7.26 (m, 5H), 5.17 (m, 2H), 4.05-3.75(m, 4H), 3.29-3.05 (m, 1H), 2.36-2.09 (m, 2H).

b) Enantiomer separation of tetrahydro-furan-3-carboxylic acid benzylester

Method Information:

Column: Chiralpak AD-PREP

Solvent: HEPTANE/ETOH/MEOH 95/2.5/2.5

Flow: 1.0 ml/min

Long onde: 210 nm

Engine: Agilent 1200 DAD Magellan

Solution EtOH

After separation of 6.347 g of racemic, 2 peaks were obtained: peak 1 at9.086 min (2.43 g, ee>99%) and peak 2 at 10.584 min (2.19 g, ee>99%).

HPLC (peak 1 or 2) Rt_(M2)=0.92 min; ESIMS: 207 [(M+H)⁺].

¹H NMR (peak 1) (400 MHz, CDCl₃): δ 7.48-7.29 (m, 5H), 5.23-5.07 (m,2H), 4.05-3.77 (m, 4H), 3.23-3.10 (m, 1H), 2.35-2.06 (m, 2H).

¹H NMR (peak 2) (400 MHz, CDCl₃): δ 7.51-7.31 (m, 5H), 5.23-5.09 (m,2H), 4.07-3.77 (m, 4H), 3.24-3.07 (m, 1H), 2.35-2.06 (m, 2H).

c) (S)-Tetrahydro-furan-3-carboxylicacid/(R)-Tetrahydro-furan-3-carboxylic acid

A solution of an enantiomerically pure tetrahydro-furan-3-carboxylicacid benzyl ester (peak 1 or peak 2, 200 mg, 0.97 mmol), Pd/C (103 mg,0.97 mmol) in EtOH (2 ml) was hydrogenated with H₂ at rt for 18 h.Filtration of the reaction mixture and concentration of the filtrateafforded the title compound as colourless oil (125 mg (Peak 1), 111 mg(Peak 2), crude).

¹H NMR (both enantiomers) (400 MHz, DMSO-d₆): δ 12.40 (br s, 1H),3.84-3.59 (m, 4H), 3.00 (m, 1H), 2.08-1.92 (m, 1H).

Example IB35-tert-Butoxycarbonylamino-1-methyl-1H-imidazole-4-carboxylic acid a)5-di(tert-Butoxycarbonyl)amino-1-methyl-1H-imidazole-4-carboxylic acidethyl ester

A solution of 5-amino-1-methyl-1H-imidazole-4-carboxylic acid ethylester (CAS registry 54147-04-5) (82 mg, 0.49 mmol) in THF (4 ml) wastreated at −50° C. with 1M LiHMDS in THF soln. (0.97 ml, 0.97 mmol) andthe reaction mixture was stirred at −50° C. for 10 min, then a solutionof Boc₂O (237 mg, 1.07 mmol) in THF (1.5 ml) was added. The reactionmixture was allowed to warm slowly to rt, then to 50° C. and to stir for15 h at 50° C. The reaction mixture was cooled to rt, diluted with EtOAcand quenched with H₂O. The organic layer was dried over Na₂SO₄,filtered, concentrated and the title compound was obtained after flashchromatography on silica gel (heptane/EtOAc, 85:15 to 0:100) as a whitesolid (140 mg, 78% yield).

UPLC Rt_(M1)=0.96 min; ESIMS: 370 [(M+H)⁺]

¹H NMR (400 MHz, CDCl₃, 298 K): δ 4.33 (q, 2H), 3.50 (s, 3H), 1.41 (s,18H), 1.35 (t, 3H).

b) 5-tert-Butoxycarbonylamino-1-methyl-1H-imidazole-4-carboxylic acid

A solution of5-di(tert-butoxycarbonyl)amino-1-methyl-1H-imidazole-4-carboxylic acidethyl ester (167 mg, 0.452 mmol) in THF (2.2 ml) and H₂O (2.2 ml) wastreated with LiOH (54.1 mg, 2.26 mmol) and the reaction mixture wasstirred at rt for 18 h, then quenched with 1M aq. HCl soln. to reach pH2 and extracted with EtOAc. The organic layer was dried over Na₂SO₄,filtered, concentrated to afford the title compound. (70 mg, 65% yield).

UPLC Rt_(M1)=0.47 min; ESIMS: 242 [(M+H)⁺]

¹H NMR (400 MHz, DMSO-d₆): δ 12.42 (br.s., 1H), 8.98 (br.s., 1H), 7.82(s, 1H), 3.45 (s, 3H), 1.41 (s, 9H).

IC) Pyrrolidinol Derivatives or Analogues

Inter- Used for Comment on mediate # Autonom name example # synthesisIC1 Methanesulfonic acid C1 2 steps from (R) -(tetrahydro- CAS104706-47-0 pyran-4-carbonyl)- pyrrolidin-3-yl ester IC2 Methanesulfonicacid C2 2 steps from (S)-(tetrahydro- CAS 100243-39-8 pyran-4-carbonyl)-pyrrolidin-3-yl ester IC3 Methanesulfonic acid C3 2 steps from(S)-1-propionyl- CAS 100243-39-8 pyrrolidin-3-yl ester IC4Methanesulfonic acid C4 2 steps from (R)-1-propionyl- CAS 2799-21-5pyrrolidin-3-yl ester

Intermediate IC1 Methanesulfonic acid(R)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yl ester a)((R)-3-Hydroxy-pyrrolidin-1-yl)-(tetrahydro-pyran-4-yl)-methanone

Oxalyl chloride (0.20 ml, 3.84 mmol) was added to a solution oftetrahydro-pyran-4-carboxylic acid (CAS registry 5337-03-1) and DMF(0.012 ml, 0.15 mmol). The reaction mixture was stirred at 3° C. for 1h. Concentration of the reaction mixture under reduced pressure (170mbar) at 40° C. (water bath) afforded the acyl intermediate as acolourless oil. The intermediate was dissolved in DCM (2 ml) and addedto a solution of (R)-pyrrolidin-3-ol hydrochloride (CAS registry104706-47-0) (190 mg, 1.54 mmol) in DCM (3 ml), cooled down to 3° C.,the formed white suspension was stirred at 3° C. for 1 h. The reactionmixture was concentrated; EtOAc was added to the residue which wasfiltered off and washed with EtOAc. Concentration and purification ofthe filtrate by flash chromatography on silica gel (DCM/DCM: MeOH (9:1),100:0 to 60:40 over 11 min) afforded the title compound as white solid(250 mg, 82% yield).

ESIMS: 200 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 4.43-4.65 (m, 1H), 3.95-4.16 (m, 2H),3.30-3.82 (m, 6H), 2.45-2.75 (m, 1H), 1.83-2.30 (m, 4H), 1.54-1.78 (m,3H).

b) Methanesulfonic acid(R)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yl ester

Under argon, methanesulfonyl chloride (CAS registry 124-63-0) (3.52 ml,45.2 mmol) was added to a solution of((R)-3-hydroxy-pyrrolidin-1-yl)-(tetrahydro-pyran-4-yl)-methanone (6 g,22.6 mmol) and Et₃N (6.30 ml, 45.2 mmol) in DCM (100 ml) at −10° C. Thesolution was stirred at 0° C. for 1 h, diluted with H₂O and DCM, theorganic layer was washed twice with H₂O and brine and dried over MgSO₄.Concentration and trituration of the resulting oil with diethyl etherafforded the title compound as an off-white solid (5.3 g, 84%).

ESIMS: 278 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 5.22-5.44 (m, 1H), 3.96-4.13 (m, 2H),3.85-3.96 (m, 1H), 3.56-3.83 (m, 3H), 3.36-3.53 (m, 2H), 3.08 (d, 3H),2.07-2.75 (m, 3H), 1.93 (m, 2H), 1.51-1.75 (m, 3H).

Intermediate IC2 Methanesulfonic acid(S)-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yl ester a)((S)-3-Hydroxy-pyrrolidin-1-yl)-(tetrahydro-pyran-4-yl)-methanone

A solution of tetrahydro-pyran-4-carbonyl chloride (CAS registry40191-32-0) (316 mg, 2.02 mmol) in DCM (2 ml) was dropwise added (0°C.<T<10° C.) to a solution of (S)-pyrrolidin-3-ol (CAS registry100243-39-8) (250 mg, 2.02 mmol) and Et₃N (0.62 ml, 4.45 mmol) in DCM (5ml). The reaction mixture was stirred at 3° C. for 1 h. The volatileswere concentrated and EtOAc was added to the residue, remaining solidwas filtered off and washed with EtOAc, concentration of the filtrateafforded the title compound as a white solid (390 mg, 97% yield).

ESIMS: 200 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 4.55 (d, 1H), 3.96-4.11 (m, 2H), 3.36-3.80(m, 6H), 2.48-2.74 (m, 1H), 1.52-2.20 (m, 7H).

b) Methanesulfonic acid(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yl ester

Under argon, methanesulfonyl chloride (CAS registry 124-63-0) (0.23 ml,2.94 mmol) was added to a solution of((S)-3-hydroxy-pyrrolidin-1-yl)-(tetrahydro-pyran-4-yl)-methanone (0.39g, 1.96 mmol) and Et₃N (0.55 ml, 3.91 mmol) in DCM (10 ml) at −10° C.The solution was stirred at 3° C. for 1 h, diluted with H₂O and DCM, theorganic layer was washed twice with H₂O and brine, then dried overMgSO₄. Concentration and trituration of the resulting oil with diethylether afforded the title compound as white solid (0.45 g, 83% yield).

ESIMS: 278 [(M+H)⁺].

¹H NMR (400 MHz, CDCl₃): δ 5.25-5.44 (m, 1H), 3.99-4.16 (m, 2H),3.85-3.95 (m, 1H), 3.56-3.83 (m, 3H), 3.37-3.54 (m, 2H), 3.08 (d, 3H),2.09-2.78 (m, 3H), 1.93 (m, 2H), 1.51-1.76 (m, 2H)

Intermediate IC3 Methanesulfonic acid (S)-1-propionyl-pyrrolidin-3-ylester a) 1-((S)-3-Hydroxy-pyrrolidin-1-yl)-propan-1-one

Propionyl chloride (CAS registry 79-03-8) (4.78 ml, 54.8 mmol) wasdropwise added over a period of 15 min to a solution of(S)-pyrrolidin-3-ol (CAS registry 100243-39-8) (4.8 g, 55.9 mmol) andEt₃N (8.74 ml, 63.1 mmol) at 5° C., the solution was allowed to warm tort and was stirred for 1 h. H₂O (10 ml) and sat. aq. NaHCO₃ soln (10 ml)were added to the solution, the organic phase was washed with brine (10ml) and a 0.25M aq. HCl soln (20 ml). The combined aqueous layers wereconcentrated and extracted with EtOAc (2×100 ml), the combined organicphases were dried over MgSO₄ and concentrated to afford the titlecompound as a pale yellow oil (4.80 g, 54% yield).

¹H NMR (400 MHz, CDCl₃): δ 4.54 (d, 1H), 3.31-3.73 (m, 3H), 3.12 (m,1H), 1.82-2.46 (m, 5H), 1.17 (t, 3H)

b) Methanesulfonic acid (S)-1-propionyl-pyrrolidin-3-yl ester

Under argon, methanesulfonyl chloride (CAS registry 124-63-0) (1.36 ml,17.46 mmol) was added over a period of 10 min to a solution of1-((S)-3-hydroxy-pyrrolidin-1-yl)-propan-1-one (2.5 g, 17.4 mmol) andEt₃N (2.43 ml, 17.4 mmol) in DCM (50 ml) at 5° C. The solution wasallowed to warm to rt and was stirred for 18 h, methanesulfonyl chloride(CAS registry 124-63-0) (1.36 ml, 17.46 mmol) and Et₃N (2.43 ml, 17.4mmol) were then added to the reaction mixture which was stirred for 48 hat rt. DCM and H₂O were added to the solution and the organic phase wasseparated through a phase separation cartridge, concentrated and thetitle compound was obtained after flash chromatography on silica gel(EtOAc/MeOH 100:0 to 95:5 over 40 min) as a colourless oil (2.7 g, 66%yield).

¹H NMR (400 MHz, CDCl₃): δ 5.19-5.42 (m, 1H), 3.48-3.99 (m, 4H), 3.06(d, 3H). 2.04-2.54 (m, 4H), 1.16 (t, 3H).

Intermediate IC4 Methanesulfonic acid (R)-1-propionyl-pyrrolidin-3-ylester a) 1-((R)-3-Hydroxy-pyrrolidin-1-yl)-propan-1-one

Propionyl chloride (CAS registry 79-03-8) (7.06 ml, 81 mmol) was added(0° C.<T<10° C.) over a period of 15 min to a suspension of(R)-pyrrolidin-3-ol (CAS registry 2799-21-5) (10 g, 81 mmol) and Et₃N(23.6 ml, 170 mmol) in DCM (150 ml) that was precooled at −10° C. Theoff-white suspension was stirred at 0° C. for 2 h, MeOH (9.82 ml, 243mmol) was added and the mixture was allowed to stir at rt for 1 h.Concentration and dilution of the residue with Et₂O (200 ml) afforded,after filtration and concentration of the filtrate, the title compoundas a yellow oil (11.2 g, 95%).

ESIMS: 144 [(M+H)⁺].

¹H NMR (400 MHz, DMSO-d₆): δ 4.81-5.04 (m, 1H), 4.15-4.38 (m, 1H),3.35-3.59 (m, 2H), 3.16-3.29 (m, 2H), 2.11-2.33 (m, 2H), 1.65-2.00 (m,2H), 0.98 (td, 3H).

b) Methanesulfonic acid (R)-1-propionyl-pyrrolidin-3-yl ester

Under argon, methanesulfonyl chloride (CAS registry 124-63-0) (0.16 ml,2.09 mmol) was dropwise added over a period of 5 min to a solution of1-((R)-3-hydroxy-pyrrolidin-1-yl)-propan-1-one (300 mg, 2.09 mmol) andEt₃N (0.29 ml, 2.09 mmol) in DCM (10 ml) at 5° C., the reaction mixturewas allowed to stir at rt for 18 h. Methanesulfonyl chloride (CASregistry 124-63-0) (0.16 ml, 2.09 mmol) and Et₃N (0.29 ml, 2.09 mmol)were then added to the reaction mixture which was stirred at rt for 48h. DCM and H₂O were added to the solution and the organic phase wasseparated through a phase separation cartridge and concentrated. Thetitle compound was obtained after flash chromatography on silica gel(EtOAc/MeOH 100:0 to 95:5 over 25 min) as a colourless oil (420 mg,86%).

¹H NMR (400 MHz, CDCl₃): δ 5.21-5.44 (m, 1H). 3.49-4.03 (m, 4H),2.98-3.12 (m, 3H), 1.97-2.54 (m, 4H), 1.18 (t, 3H).

ID) DBO Derivatives or Analogues

Inter- ID) DBO derivative or Comment mediate analogue on # StructureAutonom name synthesis ID1

3,3-Dideutero-3,4- dihydro-2H- benzo[1,4]oxazin- 6-ol 2 steps from CAS53412- 38-7, see Example Q steps a), b)

Biological Evaluation

The activity of a compound according to the present invention can beassessed by the following in vitro & in vivo methods.

Biological Assays

1 Determination of enzymatic PI3K alpha and PI3K Delta IsoformInhibition

1.1 Test of Lipid Kinase Activity

The efficacy of the compounds of examples 1-117 as PI3 kinase inhibitorscan be demonstrated as follows:

The kinase reaction is performed in a final volume of 50 μl per well ofa half area COSTAR, 96 well plate. The final concentrations of ATP andphosphatidyl inositol in the assay are 5 μM and 6 μg/mL, respectively.The reaction is started by the addition of PI3 kinase, e.g. PI3 kinaseδ.

p110δ. The components of the assay are added per well as follows:

-   -   10 μl test compound in 5% DMSO per well in columns 2-1.    -   Total activity is determined by addition 10 μl of 5% vol/vol        DMSO in the first 4 wells of column 1 and the last 4 wells of        column 12.    -   The background is determined by addition of 10 μM control        compound to the last 4 wells of column 1 and the first 4 wells        of column 12.    -   2 mL ‘Assay mix’ are prepared per plate:        -   1.912 mL of HEPES assay buffer        -   8.33 μl of 3 mM stock of ATP giving a final concentration of            5 μM per well        -   1 μl of [³³P]ATP on the activity date giving 0.05 μCi per            well        -   30 μl of 1 mg/mL PI stock giving a final concentration of 6            g/mL per well        -   5 μl of 1 M stock MgCl₂ giving a final concentration of 1 mM            per well    -   20 μl of the assay mix are added per well.    -   2 mL ‘Enzyme mix’ are prepared per plate (x* μl PI3 kinase p110β        in 2 mL of kinase buffer). The ‘Enzyme mix’ is kept on ice        during addition to the assay plates.    -   20 μl ‘Enzyme mix’ are added/well to start the reaction.    -   The plate is then incubated at room temperature for 90 minutes.    -   The reaction is terminated by the addition of 50 μl WGA-SPA bead        (wheat germ agglutinin-coated Scintillation Proximity Assay        beads) suspension per well.    -   The assay plate is sealed using TopSeal-S (heat seal for        polystyrene microplates, PerkinElmer LAS [Deutschland] GmbH,        Rodgau, Germany) and incubated at room temperature for at least        60 minutes.    -   The assay plate is then centrifuged at 1500 rpm for 2 minutes        using the Jouan bench top centrifuge (Jouan Inc., Nantes,        France).    -   The assay plate is counted using a Packard TopCount, each well        being counted for 20 seconds.    -   The volume of enzyme is dependent on the enzymatic activity of        the batch in use.

In a more preferred assay, the kinase reaction is performed in a finalvolume of 10 μl per well of a low volume non-binding CORNING, 384 wellblack plate (Cat. No. #3676). The final concentrations of ATP andphosphatidyl inositol (PI) in the assay are 1 μM and 10 μg/mL,respectively. The reaction is started by the addition of ATP.

The components of the assay are added per well as follows:

50 nl test compounds in 90% DMSO per well, in columns 1-20, 8concentrations (1/3 and 1/3.33 serial dilution step) in single.

-   -   Low control: 50 nl of 90% DMSO in half the wells of columns        23-24 (0.45% in final).    -   High control: 50 nl of reference compound (e.g. compound of        Example 7 in WO 2006/122806) in the other half of columns 23-24        (2.5 μM in final).    -   Standard: 50 nl of reference compound as just mentioned diluted        as the test compounds in columns 21-22.    -   20 mL ‘buffer’ are prepared per assay:        -   200 μl of 1M TRIS HCl pH7.5 (10 mM in final)        -   60 μl of 1M MgCl₂ (3 mM in final)        -   500 μl of 2M NaCl (50 mM in final)        -   100 μl of 10% CHAPS (0.05% in final)        -   200 μl of 100 mM DTT (1 mM in final)        -   18.94 mL of nanopure water    -   10 mL ‘PI’ are prepared per assay:        -   200 μl of 1 mg/mL 1-alpha-Phosphatidylinositol (Liver            Bovine, Avanti Polar Lipids Cat. No. 840042C MW=909.12)            prepared in 3% OctylGlucoside (10 μg/mL in final)        -   9.8 mL of ‘buffer’    -   10 mL ‘ATP’ are prepared per assay:        -   6.7 μl of 3 mM stock of ATP giving a final concentration of            1 μM per well        -   10 mL of ‘buffer’    -   2.5 mL of each PI3K construct are prepared per assay in ‘PI’        with the following final concentration:        -   10 nM PI3K alfa EMV B1075        -   25 nM beta EMV BV949        -   10 nM delta EMV BV1060        -   150 nM gamma EMV BV950    -   5 μl of ‘PI/PI3K’ are added per well.    -   5 μl ‘ATP’ are added per well to start the reaction.    -   The plates are then incubated at room temperature for 60 minutes        (alfa, beta, delta) or 120 minutes (gamma).    -   The reaction is terminated by the addition of 10 μl Kinase-Glo        (Promega Cat. No. #6714).    -   The assay plates are read after 10 minutes in Synergy 2 reader        (BioTek, Vermont USA) with an integration time of 100        milliseconds and sensitivity set to 191.    -   Output: The High control is around 60′000 counts and the Low        control is 30′000 or lower    -   This luminescence assay gives a useful Z′ ratio between 0.4 and        0.7

The Z′value is a universal measurement of the robustness of an assay. AZ′ between 0.5 and 1.0 is considered an excellent assay.

For this assay, the PI3K constructs mentioned are prepared as follows:

1.2 Generation of Gene Constructs

Two different constructs, BV 1052 and BV 1075, are used to generate thePI3 Kinase a proteins for compound screening.

PI3Kα BV-1052 p85(iSH2)-Gly Linker-p110a(D20aa)-C-Term His Tag

PCR products for the inter SH2 domain (iSH2) of the p85 subunit and forthe p110-a subunit (with a deletion of the first 20 amino acids) aregenerated and fused by overlapping PCR.

The iSH2 PCR product is generated from first strand cDNA using initiallyprimers

(SEQ ID NO: 1) gwG130-p01 (5′-CGAGAATATGATAGATTATATGAAGAAT-3′) and(SEQ ID NO: 2) gwG130-p02 (5′-TGGTTT-AATGCTGTTCATACGTTTGTCAAT-3′).

Subsequently in a secondary PCR reaction, Gateway (Invitrogen AG, Basel,Switzerland) recombination AttB1 sites and linker sequences are added atthe 5′end and 3′end of the p85 iSH2 fragment respectively, using primers

gwG130-p03 (SEQ ID NO: 3)(5′-GGGACAAGTTTGTACAAAAAAGCAGGCTACGAAGGAGATATACATAT-GCGAGAATATGATAGATTATATGAAGAAT-3′) and gwG152-p04 (SEQ ID NO: 4)(5′-TACCATAATTCCACCACCACCACCGGAAATTCCCCCTGGTTT-AATGCTGTTCATACGTTTGTCAAT-3′).

The p110-a fragment is also generated from first strand cDNA, initiallyusing primers

gwG152-p01 (SEQ ID NO: 5) (5′-CTAGTGGAATGTTTACTACCAAATGG-3′) andgwG152-p02 (SEQ ID NO: 6) (5′-GTTCAATG-CATGCTGTTTAATTGTGT-3′).

In a subsequent PCR reaction, linker sequence and a Histidine tag areadded at the 5′end and 3′end of the p110-a fragment respectively, usingprimers

gw152-p03 (SEQ ID NO: 7) (5′-GGGGGAATTTCCGGTGGTGGTGGTGGAATTATGGTAC-TAGTGGAATGTTTACTACC-AAATGGA-3′) and gwG152-p06 (SEQ ID NO: 8)(5′-AGCTCCGTGATGGTGATGGTGATGTGCTCCGTTCAATG-CATG CTGTTTAATTGTGT-3′).

The p85-iSH2/p110-a fusion protein is assembled in a third PCR reactionby the overlapping linkers at the 3′end of the iSH2 fragment and the5′end of the p110-a fragment, using the above mentioned gwG130-p03primer and a primer containing an overlapping Histidine tag and theAttB2 recombination sequences

(SEQ ID NO: 9) (5′-GGGACCACTTTGTACAAGAAAGCTGGGTTTAAGCTCCGTGATGGTGATGGTGAT-GTGCTCC-3′).

This final product is recombined in a (Invitrogen) OR reaction into thedonor vector pDONR201 to generate the ORF318 entry clone. This clone isverified by sequencing and used in a Gateway LR reaction to transfer theinsert into the Gateway adapted pBlueBac4.5 (Invitrogen) vector forgeneration of the baculovirus expression vector LR410.

PI3Kα BV-1075 p85(iSH2)-12 XGly Linker-p110a(D20aa)-C-Term His Tag

The construct for Baculovirus BV-1075 is generated by a three-partligation comprised of a p85 fragment and a p110-a fragment cloned intovector pBlueBac4.5. The p85 fragment is derived from plasmid p1661-2digested with Nhe/Spe. The p110-a fragment derived from LR410 (seeabove) as a SpeI/HindIII fragment. The cloning vector pBlueBac4.5(Invitrogen) is digested with Nhe/HindIII. This results in the constructPED 153.8

The p85 component (iSH2) is generated by PCR using ORF 318 (describedabove) as a template and one forward primer

KAC1028 (SEQ ID NO: 10) (5′-GCTAGCATGCGAGAATATGATAGATTATATGAAGAATATACC)and two reverse primers, KAC1029 (SEQ ID NO: 11)(5′-GCCTCCACCACCTCCGCCTGGTTTAATGCTGTTCATACGTTTGTC) and KAC1039(SEQ ID NO: 12) (5′-TACTAGTCCGCCTCCACCACCTCCGCCTCCACCACCTCCGCC).

The two reverse primers overlap and incorporate the 12× Gly linker andthe N-terminal sequence of the p110a gene to the SpeI site. The 12× Glylinker replaces the linker in the BV1052 construct. The PCR fragment iscloned into pCR2.1 TOPO (Invitrogen). Of the resulting clones, p1661-2is determined to be correct. This plasmid is digested with Nhe and SpeIand the resulting fragment is gel-isolated and purified for sub-cloning.The p110-a cloning fragment is generated by enzymatic digest of cloneLR410 (see above) with Spe I and HindIII. The SpeI site is in the codingregion of the p110a gene. The resulting fragment is gel-isolated andpurified for sub-cloning.

The cloning vector, pBlueBac4.5 (Invitrogen) is prepared by enzymaticdigestion with Nhe and HindIII. The cut vector is purified with Qiagen(Quiagen N.V, Venlo, Netherlands) column and then dephosphorylated withCalf Intestine alkaline phosphatase (CIP) (New England BioLabs, Ipswich,Mass.). After completion of the CIP reaction the cut vector is againcolumn purified to generate the final vector. A 3 part ligation isperformed using Roche Rapid ligase and the vendor specifications.

PI3Kβ BV-949 p85(iSH2)-Gly Linker-p110b(Full-Length)-C-Term His Tag

PCR products for the inter SH2 domain (iSH2) of the p85 subunit and forthe full-length p110-b subunit are generated and fused by overlappingPCR.

The iSH2 PCR product is generated from first strand cDNA initially usingprimers

gwG130-p01 (SEQ ID NO: 1) (5′-CGAGAATATGATAGATTATATGAAGAAT-3′) andgwG130-p02 (SEQ ID NO: 2) (5′-TGGTTT-AATGCTGTTCATACGTTTGTCAAT-3′).

Subsequently, in a secondary PCR reaction Gateway (Invitrogen)recombination AttB1 sites and linker sequences are added at the 5′endand 3′end of the p85 iSH2 fragment respectively, using primers

gwG130-p03 (SEQ ID NO: 3)(5′-GGGACAAGTTTGTACAAAAAAGCAGGCTACGAAGGAGATA-TACATATGCGAGAATATGATAGATTATATGAAGAAT-3′) and gwG130-p05 (SEQ ID NO: 13)(5′-ACTGAAGCATCCTCCTCCTCCTCCTCCTGGTTTAAT-GCTGTT CATACGTTTGTC-3′).

The p110-b fragment is also generated from first strand cDNA initiallyusing primers

gwG130-p04 (SEQ ID NO: 4)(5′-ATTAAACCAGGAGGAGGAGGAGGAGGATGCTTCAGTTTCATAA TGCC-TCCTGCT-3′)

which contains linker sequences and the 5′end of p110-b and

gwG130-p06 (SEQ ID NO: 14)(5′-AGCTCCGTGATGGTGATGGTGATGTGCTCCAGATCTGTAGTCT TT-CCGAACTGTGTG-3′)

which contains sequences of the 3′end of p110-b fused to a Histidinetag.

The p85-iSH2/p110-b fusion protein is assembled by an overlapping PCR areaction of the linkers at the 3′end of the iSH2 fragment and the 5′endof the p110-b fragment, using the above mentioned gwG130-p03 primer anda primer containing an overlapping Histidine tag and the AttB2recombination sequences

(SEQ ID NO: 15) (5′-GGGACCACTTTGTACAAGAAAGCTGGGTTT-AAGCTCCGTGATGGTGATGGTGATGTGCTCC-3′).

This final product is recombined in a Gateway (Invitrogen) OR reactioninto the donor vector pDONR201 to generate the ORF253 entry clone. Thisclone is verified by sequencing and used in a Gateway LR reaction totransfer the insert into the Gateway adapted pBlueBac4.5 (Invitrogen)vector for generation of the baculovirus expression vector LR280.

PI3Kδ BV-1060 p85(iSH2)-Gly Linker-p110d(Full-Length)-C-Term His Tag

PCR products for the inter SH2 domain (iSH2) of the p85 subunit and forthe full-length p110-d subunit are generated and fused by overlappingPCR.

The iSH2 PCR product is generated from first strand cDNA using initiallyprimers

gwG130-p01 (SEQ ID NO: 1) (5′-CGAGAATATGATAGATTATATGAAGAAT-3′) andgwG130-p02 (SEQ ID NO: 2) (5′-TGGTTT-AATGCTGTTCATACGTTTGTCAAT-3′).

Subsequently, in a secondary PCR reaction Gateway (Invitrogen)recombination AttB1 sites and linker sequences are added at the 5′endand 3′end of the p85 iSH2 fragment respectively, using primers

gwG130-p03 (SEQ ID NO: 3)(5′-GGGACAAGTTTGTACAAAAAAGCAGGCTACGAAGGAGATATACAT-ATGCGAGAATATGATAGATTATATGAAGAAT-3′) and gwG154-p04 (SEQ ID NO: 16)(5′-TCCTCCTCCTCCTCCTCCTGGTTTAATGCTGTTCATACGTTTG TC-3′).

The p110-a fragment is also generated from first strand cDNA usinginitially primers

gwG154-p01 (SEQ ID NO: 17) (5′-ATGCCCCCTGGGGTGGACTGCCCCAT-3′) andgwG154-p02 (SEQ ID NO: 18) (5′-CTACTG-CCTGTTGTCTTTGGACACGT-3′).

In a subsequent PCR reaction linker sequences and a Histidine tag isadded at the 5′end and 3′end of the p110-d fragment respectively, usingprimers

gw154-p03 (SEQ ID NO: 19)(5′-ATTAAACCAGGAGGAGGAGGAGGAGGACCCCCTGGGGTGGAC- TGCCCCATGGA-3′) andgwG154-p06 (SEQ ID NO: 20)(5′-AGCTCCGTGATGGTGAT-GGTGATGTGCT-CCCTGCCTGTTGT CTTTGGACACGTTGT-3′).

The p85-iSH2/p110-d fusion protein is assembled in a third PCR reactionby the overlapping linkers at the 3′end of the iSH2 fragment and the5′end of the p110-d fragment, using the above mentioned gwG130-p03primer and a primer containing an overlapping Histidine tag and theGateway (Invitrogen) AttB2 recombination sequences

(SEQ ID NO: 21) (5′-GGGACCACTTTGTA-CAAGAAAGCTGGGTTT-AAGCTCCGTGATGGTGATGGTGATGTGCTCC-3′).

This final product is recombined in a Gateway (Invitrogen) OR reactioninto the donor vector pDONR201 to generate the ORF319 entry clone. Thisclone is verified by sequencing and used in a Gateway LR reaction totransfer the insert into the Gateway adapted pBlueBac4.5 (Invitrogen)vector for generation of the baculovirus expression vector LR415.

PI3Kv BV-950 p110g(D144aa)-C-Term His Tag

This construct is obtained from Roger Williams lab, MRC Laboratory ofMolecular Biology, Cambridge, UK (November, 2003). Description of theconstruct in: Pacold M. E. et al. (2000) Cell 103, 931-943.

1.3 Protein Expression and Purification

Methods to generate recombinant baculovirus and protein for PI3Kisoforms: The pBlue-Bac4.5 (for a, b, and d isoforms) or pVL1393 (for g)plasmids containing the different PI3 kinase genes are co-transfectedwith BaculoGold WT genomic DNA (BD Biosciences, Franklin Lakes, N.J.,USA) using methods recommended by the vendor. Subsequently, therecombinant baculovirus obtained from the transfection isplaque-purified on Sf9 insect cells to yield several isolates expressingrecombinant protein. Positive clones are selected by anti-HIS oranti-isoform antibody western. For PI3K alpha and delta isoforms, asecondary plaque-purification is performed on the first clonal virusstocks of PI3K. Amplification of all baculovirus isolates is performedat low multiplicity of infection (moi) to generate high-titer, lowpassage stock for protein. production. The baculoviruses are designatedBV1052 (α) and BV1075 (α), BV949 (β), BV1060 (δ) and BV950 (γ).

Protein production involves infection (passage 3 or lower) of suspendedTn5 (Trichoplusia ni) or TiniPro (Expression Systems, LLC, Woodland,Calif., USA) cells in protein-free media at moi of 2-10 for 39-48 hoursin 2 I glass Erlenmyer flasks (110 rpm) or wave-bioreactors (22-25 rpm).Initially, 10 l working volume wave-bioreactors are seeded at a densityof 3e5 cells/mL at half capacity (5 L). The reactor is rocked at 15 rpmduring the cell growth phase for 72 hours, supplemented with 5% oxygenmixed with air (0.2 l per minute). Immediately prior to infection, thewave-reactor cultures are analyzed for density, viability and diluted toapproximately 1.5e6 cell/mL. 100-500 mL of high titer, low passage virusis added following 2-4 hours of additional culture. Oxygen is increasedto 35% for the 39-48 hour infection period and rocking platform rpmincreased to 25. During infection, cells are monitored by Vicellviability analyzer (Beckman Coulter, Inc, Fullerton, Calif., USA)bioprocess for viability, diameter and density. Nova Bioanalyzer (NOVABiomedical Corp., Waltham, Mass., USA) readings of various parametersand metabolites (pH, O₂ saturation, glucose, etc.) are taken every 12-18hours until harvest. The wave-bioreactor cells are collected within 40hours post infection. Cells are collected by centrifugation (4 degreesC. at 1500 rpm), and subsequently maintained on ice during pooling ofpellets for lysis and purification. Pellet pools are made with smallamounts of cold, un-supplemented Grace's media (w/o proteaseinhibitors).

PI3K alpha Purification Protocol for HTS (BV1052)

PI3K alpha is purified in three chromatographic steps: immobilized metalaffinity chromatography on a Ni Sepharose resin (GE Healthcare,belonging to General Electric Company, Fairfield, Conn., USA), gelfiltration utilizing a Superdex 200 26/60 column (GE Healthcare), andfinally a cation exchange step on a SP-XL column (GE Healthcare). Allbuffers are chilled to 4° C. and lysis is performed chilled on ice.Column fractionation is performed rapidly at room temperature.

Typically frozen insect cells are lysed in a hypertonic lysis buffer andapplied to a prepared IMAC column. The resin is washed with 3-5 columnvolumes of lysis buffer, followed by 3-5 column volumes wash buffercontaining 45 mM imidazole, and the target protein is then eluted with abuffer containing 250 mM imidazole. Fractions are analyzed by Coomassiestained SDS-PAGE gels, and fractions containing target protein arepooled and applied to a prepared GFC column. Fractions from the GFCcolumn are analyzed by Coomassie stained SDS-PAGE gels, and fractionscontaining target protein are pooled. The pool from the GFC column isdiluted into a low salt buffer and applied to a prepared SP-XL column.The column is washed with low salt buffer until a stable A280 baselineabsorbance is achieved, and eluted using a 20 column volume gradientfrom 0 mM NaCl to 500 mM NaCl. Again, fractions from the SP-XL columnare analyzed by Coomassie stained SDS-PAGE gels, and fractionscontaining the target protein are pooled. The final pool is dialyzedinto a storage buffer containing 50% glycerol and stored at −20° C. Thefinal pool is assayed for activity in a phosphoinosititol kinase assay.

PI3K beta Purification Protocol for HTS (BV949)

PI3K beta is purified in two chromatographic steps: immobilized metalaffinity chromatography (IMAC) on a Ni Sepharose resin (GE Healthcare)and gel filtration (GFC) utilizing a Superdex 200 26/60 column (GEHealthcare). All buffers are chilled to 4° C. and lysis is performedchilled on ice. Column fractionation is performed rapidly at roomtemperature.

Typically frozen insect cells are lysed in a hypertonic lysis buffer andapplied to a prepared IMAC column. The resin is washed with 3-5 columnvolumes of lysis buffer, followed by 3-5 column volumes wash buffercontaining 45 mM imidazole, and the target protein is then eluted with abuffer containing 250 mM imidazole. Fractions are analyzed by Coomassiestained SDS-PAGE gels, and fractions containing target protein arepooled and applied to a prepared GFC column. Fractions from the GFCcolumn are analyzed by Coomassie stained SDS-PAGE gels, and fractionscontaining target protein are pooled. The final pool is dialyzed into astorage buffer containing 50% glycerol and stored at −20° C. The finalpool is assayed for activity in the phosphoinostitol kinase assay.

PI3K gamma Purification Protocol for HTS (BV950)

PI3K gamma is purified in two chromatographic steps: immobilized metalaffinity chromatography (IMAC) on a Ni Sepharose resin (GE Healthcare)and gel filtration (GFC) utilizing a Superdex 200 26/60 column (GEHealthcare). All buffers are chilled to 4° C. and lysis is performedchilled on ice. Column fractionation is performed rapidly at roomtemperature. Typically frozen insect cells are lysed in a hypertoniclysis buffer and applied to a prepared IMAC column. The resin is washedwith 3-5 column volumes of lysis buffer, followed by 3-5 column volumeswash buffer containing 45 mM imidazole, and the target protein is theneluted with a buffer containing 250 mM imidazole. Fractions are analyzedby Coomassie stained SDS-PAGE gels, and fractions containing targetprotein are pooled and applied to a prepared GFC column. Fractions fromthe GFC column are analyzed by Coomassie stained SDS-PAGE gels, andfractions containing target protein are pooled. The final pool isdialyzed into a storage buffer containing 50% glycerol and stored at−20° C. The final pool is assayed for activity in the phosphoinostitolkinase assay.

PI3K delta Purification Protocol for HTS (BV1060)

PI3K delta is purified in three chromatographic steps: immobilized metalaffinity chromatography on a Ni Sepharose resin (GE Healthcare), gelfiltration utilizing a Superdex 200 26/60 column (GE Healthcare), andfinally a anion exchange step on a Q-HP column (GE Healthcare). Allbuffers are chilled to 4° C. and lysis is performed chilled on ice.Column fractionation is performed rapidly at room temperature. Typicallyfrozen insect cells are lysed in a hypertonic lysis buffer and appliedto a prepared IMAC column. The resin is washed with 3-5 column volumesof lysis buffer, followed by 3-5 column volumes wash buffer containing45 mM imidazole, and the target protein is then eluted with a buffercontaining 250 mM imidazole. Fractions are analyzed by Coomassie stainedSDS-PAGE gels, and fractions containing the target protein are pooledand applied to a prepared GFC column. Fractions from the GFC column areanalyzed by Coomassie stained SDS-PAGE gels, and fractions containingthe target protein are pooled. The pool from the GFC column is dilutedinto a low salt buffer and applied to a prepared Q-HP column. The columnis washed with low salt buffer until a stable A280 baseline absorbanceis achieved, and eluted using a 20 column volume gradient from 0 mM NaClto 500 mM NaCl. Again, fractions from the Q-HP column are analyzed byCoomassie stained SDS-PAGE gels, and fractions containing the targetprotein are pooled. The final pool is dialyzed into a storage buffercontaining 50% glycerol and stored at −20° C. The final pool is assayedfor activity in the phosphoinostitol kinase assay.

IC₅₀ is determined by a four parameter curve fitting routine that comesalong with “excel fit”. A four parameter logistic equation is used tocalculate IC₅₀ values (IDBS XLfit) of the percentage inhibition of eachcompound at 8 concentrations (usually 10, 3.0, 1.0, 0.3, 0.1, 0.030,0.010 and 0.003 μM). Alternatively, IC₅₀ values are calculated usingidbsXLfit model 204, which is a 4 parameter logistic model.

Yet alternatively, for an ATP depletion assay, compounds of the formulaI to be tested are dissolved in DMSO and directly distributed into awhite 384-well plate at 0.5 μl per well. To start the reaction, 10 μl of10 nM PI3 kinase and 5 μg/mL 1-alpha-phosphatidylinositol (PI) are addedinto each well followed by 10 μl of 2 μM ATP. The reaction is performeduntil approx 50% of the ATP is depleted, and then stopped by theaddition of 20 μl of Kinase-Glo solution (Promega Corp., Madison, Wis.,USA). The stopped reaction is incubated for 5 minutes and the remainingATP is then detected via luminescence. IC₅₀ values are then determined.

In one embodiment of the present invention, the PI3K inhibitor, whereinsaid inhibitor has an inhibitory action on the PI3K isoform delta,wherein the range of activity, expressed as IC₅₀, in the enzymatic PI3Kdelta assay is from is between 1 nM and 500 nM.

In another embodiment of the present invention, the PI3K inhibitor,wherein said inhibitor has an inhibitory action on the PI3K isoformdelta, wherein the range of activity, expressed as IC₅₀, in theenzymatic PI3K delta assay is from is between 1 nM and 100 nM.

In another embodiment of the present invention, the PI3K inhibitor,wherein said inhibitor has an inhibitory action on the PI3K isoformdelta, wherein the range of activity, expressed as IC₅₀, in theenzymatic PI3K delta assay is from is between 0.5 nM and 10 nM.

In one embodiment of the present invention, the PI3K inhibitor, whereinsaid inhibitor has an inhibitory action on the PI3K isoform delta,wherein the range of activity, expressed as IC₅₀, in the cellular PI3Kdelta assay is from is between 1 nM and 1000 nM.

In another embodiment of the present invention, the PI3K inhibitor,wherein said inhibitor has an inhibitory action on the PI3K isoformdelta, wherein the range of activity, expressed as IC₅₀, in the cellularPI3K delta assay is from is between 1 nM and 500 nM.

In another embodiment of the present invention, the PI3K inhibitor,wherein said inhibitor has an inhibitory action on the PI3K isoformdelta where the inhibitor shows a selectivity for the PI3K isoform deltaover one or more of the other isoforms wherein this selectivity is atleast 10 fold.

In another embodiment of the present invention, the PI3K inhibitor,wherein said inhibitor has an inhibitory action on the PI3K isoformdelta where the inhibitor shows a selectivity for the PI3K isoform deltaover one or more of the other isoforms wherein this selectivity is atleast 20 fold.

In another embodiment of the present invention, the PI3K inhibitor,wherein said inhibitor has an inhibitory action on the PI3K isoformdelta where the inhibitor shows a selectivity for the PI3K isoform deltaover the different paralogs PI3K α and □β, wherein this selectivity isat least 10 fold.

In another embodiment of the present invention, the PI3K inhibitor,wherein said inhibitor has an inhibitory action on the PI3K isoformdelta where the inhibitor shows a selectivity for the PI3K isoform deltaover the different paralogs PI3K α and □β, wherein this selectivity isat least 20 fold.

In another embodiment of the present invention, the PI3K inhibitor,wherein said inhibitor has an inhibitory action on the PI3K isoformdelta, wherein the range of activity, expressed as IC₅₀, in the cellularPI3K delta assay is from is between 1 nM and 500 nM and wherein saidinhibitor has an inhibitory action on the PI3K isoform delta where theinhibitor shows a selectivity for the PI3K isoform delta over thedifferent paralogs PI3K α and □β, wherein this selectivity is at least10 fold.

In another embodiment of the present invention, the PI3K inhibitor,wherein said inhibitor has an inhibitory action on the PI3K isoformdelta, wherein the range of activity, expressed as IC₅₀, in the cellularPI3K delta assay is from is between 1 nM and 500 nM and wherein saidinhibitor has an inhibitory action on the PI3K isoform delta where theinhibitor shows a selectivity for the PI3K isoform delta over thedifferent paralogs PI3K α and □β, wherein this selectivity is at least20 fold.

2. Cellular Assays

2.1 Phosphoinositide-3 Kinase (PI3K)-Mediated Akt 1/2 (S473)Phosphorylation in Rat-1 Cells

Rat-1 cells stably overexpressing a myristoylated form of the catalyticsubunit of human phosphoinositide-3 kinase (PI3K) alpha, beta or deltawere plated in 384-well plates at a density of 7500 (PI3K alpha), 6200(PI3K beta), or 4000 (PI3K delta) cells in 30 ul complete growth medium(Dulbecco's modified Eagle's medium (DMEM high glucose) supplementedwith 10% (v/v) fetal bovine serum, 1% (v/v) MEM non essential aminoacids, 10 mM HEPES, 2 mM L-glutamine, 10 μg/mL puromycin and 1% (v/v)Penicillin/Streptomycin) and were incubated at 37% C/5% CO₂/95% humidityfor 24 h. Compounds were diluted in 384-well compound plates to obtain8-point serial dilutions for 40 test compounds in 90% DMSO, as well as 4reference compounds plus 16 high controls and 16 low (inhibited)controls. Predilution plates were prepared by dispensing pipetting 250nl of compound solutions into 384-well polypropylen plates using aHummingwell nanoliter dispensor. Compounds were prediluted by theaddition of 49.75 ul complete growth medium. 10 ul of predilutedcompound solution were transferred to the cell plate using a 384-wellpipettor, resulting in a final DMSO concentration of 0.11%. Cells wereincubated for 1 h at 37% C/5% CO₂/95% humidity. The supernatant wasremoved, the cells were lysed in 20 ul of lysis buffer for AlphaScreen®SureFire® detection.

For detection of p-AKT(Ser473), the SureFire® p-Akt 1/2 (Ser473) AssayKit (PerkinElmer, U.S.A.) was used. 5 ul of cell lysate was transferredto 384-well low volume Proxiplates for detection using a 384-wellpipettor. Addition of AlphaScreen® SureFire® reagents was done accordingto the manufacturer's protocol. First, 5 ul of reaction buffer plusactivation buffer mix containing AlphaScreen® acceptor beads was added,the plate was sealed, and incubated on a plate shaker for 2 hours atroom temperature. Second, 2 ul of dilution buffer containingAlphaScreen® donor beads was added, and the plate was incubated on plateshaker as above for a further 2 hours. The plate was read on anAlphaScreen® compatible plate reader, using standard AlphaScreen®settings.

2.2 Determination of Murine B Cell Activation

PI3Kδ has been recognized to modulate B cell function when cells arestimulated through the B cell receptor (BCR) (Okkenhaug et al. Science297:1031 (2002). For assessing the inhibitory property of compounds on Bcell activation, the upregulation of activation markers CD86 and CD69 onmurine B cells derived from mouse spleen antibody is measured afterstimulation with anti-IgM. CD69 is a well known activation marker for Band T cells (Sancho et al. Trends Immunol. 26:136 (2005). CD86 (alsoknown as B7-2) is primarily expressed on antigen-presenting cells,including B cells. Resting B cells express CD86 at low levels, butupregulate it following stimulation of e.g. the BCR or IL-4 receptor.CD86 on a B cell interacts with CD28 on T cells. This interaction isrequired for optimal T cell activation and for the generation of anoptimal IgG1 response (Carreno et al. Annu Rev Immunol. 20:29 (2002)).

Spleens from Balb/c mice are collected, splenocytes are isolated andwashed twice with RPMI containing 10% foetal bovine serum (FBS), 10 mMHEPES, 100 Units/mL penicilline/streptomycine. RPMI supplemented in thisway is subsequently referred to as medium. The cells are adjusted to2.5×10⁶ cells/mL in medium and 200 μl cell suspension (5×10⁶ cells) areadded to the appropriate wells of 96 well plates.

Then the cells are stimulated by adding 50 μl anti-IgM mAb in medium(final concentration: 30 μg/mL). After incubation for 24 hours at 37°C., the cells are stained with the following antibody cocktails:anti-mouse CD86-FITC, anti-mouse CD69-PerCP-Cy5.5, anti-mouse CD19-PerCPfor the assessment of B cells, and anti-mouse CD3-FITC, anti-mouseCD69-PE for the assessment of T cells (2 μl of each antibody/well).After one hour at room temperature (rt) in the dark the cells aretransferred to 96 Deepwell plates. The cells are washed once with 1 mLPBS containing 2% FBS and after re-suspension in 200 μl the samples areanalyzed on a FACS Calibur flow cytometer. Lymphocytes are gated in theFSC/SSC dot plot according to size and granularity and further analyzedfor expression of CD19, CD3 and activation markers (CD86, CD69). Dataare calculated from dot blots as percentage of cells positively stainedfor activation markers within the CD19+ or CD3+ population using BDCellQest Software.

For assessing the inhibitory property of compounds, compounds are firstdissolved and diluted in DMSO followed by a 1:50 dilution in medium.Splenocytes from Balb/c mice are isolated, re-suspended and transferredto 96 well plates as described above (200 μl/well). The dilutedcompounds or solvent are added to the plates (25 μl) and incubated at37° C. for 1 hour. Then the cultures are stimulated with 25 μl anti-IgMmAb/well (final concentration 30 μg/mL) for 24 hours at 37° C. andstained with anti-mouse CD86-FITC and anti-mouse CD19-PerCP (2 μl ofeach antibody/well). CD86 expression on CD19 positive B cells isquantified by flow cytometry as described above.

2.3 Determination of Rat B Cell Activation

PI3Kδ has been recognized to modulate B cell function when cells arestimulated through the B cell receptor (BCR) (Okkenhaug et al. Science297:1031 (2002). For assessing the inhibitory property of compounds on Bcell activation, the upregulation of activation markers CD86 on rat Bcells derived from whole blood is measured after stimulation withanti-IgM and recombinant IL-4. The CD86 molecule (also known as B7-2) isprimarily expressed on antigen-presenting cells, including B cells.Resting B cells express CD86 at low levels, but upregulate it followingstimulation of e.g. the BCR or IL-4 receptor. CD86 on a B cell interactswith CD28 on T cells. This interaction is required for optimal T cellactivation and for the generation of an optimal IgG1 response (Carrenoet al. Annu Rev Immunol. 20:29 (2002)).

Collection of Rat Blood

Whole blood was collected from the abdominal aorta adult male Lewis rats(LEW/HanHsd) oby using a 10 ml syringe with hypodermic needle pre-coatedwith sodium heparin. Blood was transferred into 50 ml Falcon tubes andthe anticoagulant concentration was adjusted to 100 U/ml.

Stimulation of Rat B Cells and Treatment with Specific Inhibitor

For assessment of the in vitro effects of immunosuppressive drugs,heparinized blood was prediluted to 50% with medium. As medium servedDMEM high glucose (Animed cat#1-26F01-I) supplemented with 100 U/mlpenicillin, 100 mg/ml streptomycin, 2 mM L-glutamin, 50 mg/ml dextran 40and 5% fetal calf serum (FCS, Fetaclone I, Gibco #10270-106). Then, 190μl prediluted blood was spiked with 10 μl of pre-diluted test compoundin 96 well U-bottomed microtiter plates (Nunc) resulting in a 3-foldserial dilution with a concentration range from 20 to 0.0003 μM. Controlwells were pretreated with DMSO to obtain a final concentration of 0.5%DMSO. Cultures were set up in duplicates, mixed well by agitation on aplate shaker (Heidolph Titramax 101; 30 sec, speed 900), pipetting upand down and agitated on the plate shaker again. Cultures were incubatedat 37° C., 5% CO₂ for 1 hr. Then, 20 μl of polyclonal goat anti-rat IgMAb (Serotec, cat#302001) and 10 μl of diluted recombinant rIL-4(Immunotools #340085) were added to obtain final concentrations of 30μg/ml and 5 ng/ml, respectively. Plates were mixed by agitation on aplate shaker as above and incubated for 24 hrs at 37° C., 5% CO₂.

Determination of B Cell Activation by Flow Cytometry

After incubation, 15 μl of a 25 mM EDTA solution was added per well andshaken for 15 min to detach adherent cells. For analysis of surfaceactivation markers, samples were then stained with PE-Cy5-labeledanti-ratCD45RA (BD cat#557015) to allow gating on B cells in FACSanalysis. In addition, samples were stained with PE-labeled anti-ratCD86 (BD cat#551396). All staining procedures were performed at rt for30 min in the dark. After incubation, samples were transferred to96-deep well V-bottomed microtiter plates (Corning #396096) containing 2ml/well of BD Lysing Solution (BD #349202). After lysis of erythrocytessamples were washed with 2 ml of CellWASH (BD #349524). Data wasacquired on an LSRII or FACScalibur flow cytometer (BD Biosciences)using Cellquest Plus or DIVA (version 6.1.1) software, respectively.Lymphocytes were gated in the FSC/SSC dot blot according to size andgranularity and further analyzed for expression of CD45RA and activationmarkers. Data were calculated from dot blots or histograms as percentageof cells positively stained for activation markers within the CD45RA+population.

Statistical Evaluation

The percentage inhibition of B cell activation after exposure to drugwas calculated by the following formula:

${\%\mspace{14mu}{Inhibition}} = {100 \times \frac{{{stimulation}\mspace{14mu}{without}\mspace{14mu}{drug}} - {{stimulation}\mspace{14mu}{with}\mspace{14mu}{drug}}}{{{stimulation}\mspace{14mu}{without}\mspace{14mu}{drug}} - {unstimulated}}}$

ORIGIN 7 software (OriginLab Corporation, Northampton, Mass.) was usedfor non-linear regression curve fitting. The drug concentrationresulting in 50% inhibition (IC₅₀) was obtained by fitting the Hillequation to inhibition data.

2.4 Determination of TLR9-Induced IL-6 in Mouse Splenocytes

Preparation of Single Cell Suspension from Mouse Spleen

Spleens were dissected from C57BL/6 mice immediately followingeuthanasia. Excess fat was trimmed from the spleens prior to mashing thespleen through a 0.4 μM cell strainer using a plunger from a 5 mlsyringe. A single cell suspension was prepared and the volume wasadjusted to 15 ml in a 50 ml Falcon tube using cold PBS. Cells werecentrifuged at 1500 rpm for 5 minutes at 4° C. degrees prior to removalof supernatant and re-suspension in 5 ml of red blood cell lysis bufferper spleen and incubation for 5 minutes at room temperature. Ice coldPBS (30 ml) was added to the cells prior to centrifugation at 1500 rpmfor 5 minutes at 4° C. The supernatant was removed and the cells werewashed twice with 40 ml of murine splenocyte culture media (MSCM). MSCMconsisted of RPMI supplemented with 100 units/ml Penicillin and 100μg/ml Streptomycin, 1×nonessential amino acids, 1 mM Sodium Pyruvate,0.05 mM β-mercaptoethanol, and 10% heatinactivated Fetal Bovine Serum(FBS). Cells were re-suspended in 10-20 ml of MSCM and counted using aCountess cell counter. Approximately 60×10⁶ splenocytes were obtainedfrom a single C57BL/6 mouse spleen.

Stimulation of Murine Splenocytes and Treatment with Specific Inhibitor

Splenocytes were plated at a final density of 2×10⁵ cells/well in avolume of 100 μl in 96 well flat bottomed plates and incubated in ahumidified 37° C. incubator for 2-4 hours. Afterwards, compounds to betested were dispensed using an automated liquid handling machine usingpreviously prepared compound stock plates. Stock plates consisted ofcompounds (in 90%/10% DMSO/ddH₂O) arrayed in 8-10 point using 2- or3-fold dilutions. The liquid handling machine dispensed 1 μl of eachdilution from the previously prepared compound source plate into theappropriate destination well in the 96-well plate. The final startingconcentration of the compounds in the cell culture was 10 μM. The finalconcentration of DMSO in the cell cultures was 0.5%. Cells wereincubated with compounds for 1 hour prior to addition of TLR ligand.Then, a 10× EC₈₀ concentration of CpG1826 was added in a volume of 20 μl(for a final culture volume of 200 μl) whereupon cultures were incubatedovernight in a humidified 37° C. incubator.

Determination of Interleukin-6 by ELISA

After overnight culture, plates were centrifugated at 2000 rpm for 5minutes at room temperature. Subsequently 150 μl of each culture wastransferred to 96-well V-bottomed plates and IL-6 levels were measuredusing commercially available mouse IL-6 sandwich ELISA kit. Briefly,plates were coated overnight with the capture antibody prior to blockingfor 1 hour with PBS/0.1% BSA. Samples and standards were added in avolume of 50 μl and the plate was incubated for 2 hours at roomtemperature. After removal of the standards/samples, the plate waswashed using PBS/0.05% Tween prior to addition of 50 μl of thebiotinylated detection antibody whereupon the plate was incubated for 2hours at room temperature with agitation. Plates were washed again priorto addition of 50 μl streptavidin-horseradish peroxidase per well for 20minutes. Following additional plate washes 50 μl TMB substrate was addedto each well and plates were incubated for 20 minutes prior addition of25 μl/well stop solution. IL-6 levels were measured using a SpectraMax190 Plate Reader (450 nm) and analyzed using SoftMax Pro and GraphPadPrism software.

2.5 Determination of TLR9-Induced IFNalpha in Human Peripheral BloodMononuclear Cells (PBMC)

Preparation of PBMC from Fresh Human Blood

Human blood (ca. 75 ml) was collected in 10 S-Monovette tubes containingHeparin (S-Monovette 7.5 mL NH Heparin 16 IU/mL blood; Starstedt).Leucosep™ tubes (30 mL #227290; Greiner Bio-one) were prepared byaddition of 15 ml lymphocyte separation medium LSM1077™ per tube(#J15-004; PAA Laboratories) and centrifugation for 30 sec at 1000 g.Some 25 ml blood was transferred to Leucosep™ tubes following dilutionwith equal parts of PBS (without Ca2+/Mg2+; #14190-094). Samples werecentrifuged at 800 g for 20 min at 22° C. using an Eppendorf 5810Rcentrifuge without brake. The PBMC layer was carefully removed fromplasma:separation medium interface and transferred into clean 50 mltube. Cells were washed once by addition of PBS (up to 45 ml) andcentrifuged (1400 rpm, 10 min at 22° C.) with brake (set at speed 9)using an Eppendorf 5810R. Pelleted cells were carefully resuspended inMedia (RPMI 1640+GlutaMAX-I, 0.05 mM 2-mercaptoethanol, 10 mM HEPES and5% v/v FCS) and samples pooled. The medium components 2-mercaptoethanol(#31350-010; 50 mM), Hepes (#15630-056, 1M) and RPMI 1640(1×)+GlutaMAX-I (#61870-010) were obtained from Gibco. FCS (#2-01F36-1)was obtained from Amimed. The PBMC were counted using a Countess®Automated cell counter (sample was pre-diluted 1:10 in Media, prior tothe addition of equal volume (10 μl) of Trypan Blue). Cells were dilutedto 4×10⁶ cells/ml and seeded in 384-well plates (#353962; BectonDickinson AG) to give a final volume of 25 μl (i.e. 1×10⁵ cells/well).

Stimulation of PBMC and Treatment with Specific Inhibitor

Compounds were pre-diluted in 100% v/v DMSO (#41640-100 mL;Sigma-Aldrich), followed by transfer in Media (to achieve a final DMSOconcentration of 0.25%). Cells were treated with appropriate compounddilution (5 μl) or vehicle control (5 μl) and incubated for 30 min at37° C. in a humidified incubator in air with 5% (v/v) CO₂. Cells werestimulated with CpG2216 (0.3 μM; #tlrl-hodna; Invivogen) or vehiclecontrol (10 μl/well) and incubated for 20 h. Plates were brieflycentrifuged (200×g for 2 min at 22° C.) and supernatant samples (30 μl)removed for quantification of IFNα levels.

Quantification of IFNα Using AlphaLisa Technology

For quantification of IFNalpha the human interferon AlphaLISA Kit(#AL264F) from PerkinElmer was used. An antibody mix containinganti-IFNα acceptor beads (5 μg/ml final) and biotinylated antibodyanti-IFNα (0.5 nM final) is prepared fresh and dispensed (5 μl) into384-well Optiplates™ (#6007299; PerkinElmer). Dilution of known IFNαstandards (human IFNα B (2b)) were prepared and together with cellsupernatants (5 μl) were added to plates above. Plates were brieflycentrifuged (pulse at 200 g), covered with adhesive sealing film,vortexed and incubated 1 h at room temperature in the dark.Streptavidin-coated donor beads (20 μg/ml final) was prepared and addedto each well (5 μl) in a dark lit area (light sensitive mix). Plateswere incubated 30 min at room temperature (Pates must not be centrifugedor covered). After incubation, the plates were read with an EnVision™multiplate reader equipped with the ALPHA option using the instrument'sown “AlphaScreen standard settings” (e.g. total measurement time: 550ms, Laser 680 nm excitation time: 180 ms, mirror: D640 as, emissionfilter: M570w, center wavelength 570 nm, bandwidth 100 nm, transmittance75%). Data were collected for analysis and quantification of IFNαlevels.

Data Evaluation and Analysis

Data were analysed using Excel XL fit 4.0 (Microsoft) with XLfit add-in(IDBS; version 4.3.2). Specific IFNα concentrations were determinedfollowing extrapolation to standard curves using human IFNα B (2b).Individual IC₅₀ values of compounds were determined by nonlinearregression after fitting of curves to the experimental data.

3 Determination of Antibody Production to Sheep Red Blood Cells (SRBC).

In brief, OFA rats were injected i.v. with sheep erythrocytes on d0 andtreated orally on four consecutive days (d0 to d3) with the compoundsunder investigation. Spleen cell suspensions were prepared on d4 andlymphocytes were plated onto soft agar in presence of indicator cells(SRBC) and complement. Lysis of the indicator cells due to secretion ofSRBC-specific antibody (predominantly of the IgM subclass) and presenceof complement yielded plaques. The number of plaques per plate werecounted and expressed as number of plaques per spleen.

Immunization:

Groups of five female OFA rats were immunized on day 0 with 2×10⁸/mlSRBC (obtained from Laboratory Animal Services LAS, Novartis Pharma AG)in a volume of 0.5 ml per rat by i.v. injection.

Compound Treatment:

Animals were treated with compound suspended in 0.5% CMC, 0.5% Tween80in for 4 consecutive days (days 0, 1, 2 and 3) starting on the day ofimmunization. Compound was administered orally twice daily with 12 hoursintervalls between doses in an application volume of 5 ml/kg bodyweight.

Preparation of Spleen Cell Suspensions:

On day 4, animals were euthanized with CO₂. Spleens were removed,weighed, and deposited in plastic tubes containing 10 ml of cold (4° C.)Hank's balanced salt solution (HBSS; Gibco, pH 7.3, containing 1 mgPhenol red/100 ml) for each rat spleen. Spleens were homogenized with aglass potter, left on ice for 5 minutes and 1 ml supernatant wastransferred into a new tube. Cells were washed once in 4 ml HBSS thensupernatants were discarded and pellets re-suspended in 1 ml of HBSS.Lymphocyte numbers per spleen were determined by automated cell counterand spleen cell suspensions were adjusted to a cell concentration of30×10⁶/ml.

Plaque Forming Assay:

Soft agar petri dishes were prepared with 0.7% agarose (SERVA) in HBSS.

In addition, one ml of 0.7% agarose was prepared in plastic tubes andkept at 48° C. in a water bath. Some 50 μl of a 30×10⁶/ml spleen cellsuspension and 50 μl of SRBC at 40×10⁸/ml were added, mixed rapidly(Vortex) and poured onto the prepared agarose dishes. Petri dishes wereslightly tilted to achieve even distribution of cell mixture on agaroselayer. The dishes were left at room temperature for 15 minutes and werethen incubated at 37° C. for 60 minutes. Then, 1.4 ml guinea pigcomplement (Harlan; 10%) was added and the incubation continued foranother 60 minutes at 37° C. SRBC-specific antibodies released by theplated-out B cells bound to the antigen (SRBC) in their vicinity. Theseantigen-antibody complexes activated complement and led to the lysis ofthe SRBC leaving a bright spot (plaque) within the red erythrocytelayer. Plaques were counted with a microscope.

The following formula for determination of inhibition of plaqueformation was used:% Inhibition=C*100/V−100

with: V=mean number of plaques/spleen for vehicle group; C=mean numberof plaques/spleen for compound treated group

REFERENCES

-   N. K. Jerne & A. A. Nordin (1963) Plaque formation in agar by single    antibody-producing cells. Science 140:405.-   N. K. Jerne, A. A. Nordin & C. Henry (1963) The agar plaque    technique for recognizing antibody-producing cells. In: “Cell Bound    Antibodies”, B. Amos & H. Koprowski, Eds., Wistar Inst. Press,    Philadelphia pp. 109-125.

Biological Data

Enzymatic Assay

Example PI3K alpha (uM) PI3K delta (uM) A1 0.322 0.006 A2 0.047 0.006 A30.313 0.003 A4 >9.1 A5 4.663 0.037 A6 0.377 0.009 A7 1.915 0.031 A85.928 0.04 A9 0.410 0.014 A10 0.220 0.018 A11 2.279 0.069 A12 0.1820.003 A13 0.292 0.005 A14 >9.1 5.399 A15 4.892 0.184 A16 >9.1 0.323 A170.104 0.014 A18 0.895 0.011 A19 7.547 0.38 A20 >9.1 A21 8.429 A22 0.7570.021 A23 1.573 0.17 A24 6.878 0.317 A25 5.755 A26 0.152 0.022 A27 >9.1A28 0.531 0.016 A29 2.730 0.042 A30 0.260 0.031 A31 6.022 0.088 A323.195 0.074 A33 1.702 0.085 A34 0.773 0.009 A35 5.589 1.419 A36 1.2690.078 A37 0.370 0.078 A38 0.343 0.064 A39 0.071 0.009 A40 5.361 0.462A41 2.794 0.296 A42 1.141 0.03 A43 4.689 0.054 B1 0.390 0.014 B2 1.6410.026 B3 >9.1 B4 1.234 0.083 B5 0.236 0.025 B6 2.242 0.11 B7 3.544 0.114B8 0.188 0.025 B9 1.770 0.078 B10 0.870 0.035 B11 0.823 0.014 B12 0.3650.011 B13 0.862 0.062 B14 0.244 0.006 B15 2.647 0.031 B16 4.117 0.037B17 2.087 0.023 B18 0.723 0.012 B19 1.158 0.033 B20 0.339 0.047 B210.666 0.013 B22 0.589 0.005 B23 1.004 0.028 B24 0.212 0.008 B25 0.6650.04 B26 0.281 <0.009 B27 2.507 0.055 B28 0.788 0.012 B29 0.558 0.041B30 0.216 0.0155 B31 0.977 0.003 B32 0.369 0.036 B33 0.610 <0.009 B340.795 0.046 B35 1.387 0.027 B36 0.801 <0.003 B37 1.104 0.103 B38 0.1660.003 B39 0.176 0.011 B40 0.249 0.012 B41 0.421 0.025 B42 0.502 0.033B43 0.514 0.057 B44 0.071 0.022 B45 0.574 0.0745 B46 2.076 0.2265 B470.241 0.0106667 B48 0.109 0.007 B49 0.450 0.007 B50 0.933 0.013 B511.026 0.01 B52 0.556 <0.003 B53 0.343 0.0045 B54 0.756 0.011 B55 3.8500.065 B56 1.415 0.023 B57 2.270 0.095 B58 0.366 0.004 B59 0.461 0.017B60 0.463 0.011 B61 0.732 0.037 B62 0.496 0.041 B63 1.518 0.068 B640.193 0.032 B65 0.795 0.035 B66 0.446 0.011 B67 0.804 0.025 B68 0.8400.039 B69 0.189 0.005 B70 0.134 0.003 B71 0.360 0.0055 B72 2.057 0.032B73 1.771 0.03 B74 0.054 0.009 B75 0.211 0.011 B76 0.399 0.061 B77 0.8970.032 B78 0.243 0.023 B79 0.661 0.003 B80 0.295 0.004 B81 0.482 0.081B82 0.353 0.066 B83 0.214 0.029 B84 0.346 0.08 B85 B86 6.626 0.061 B877.092 0.029 B88 0.234 0.008 B89 0.133 0.009 B90 1.456 0.011 B91 0.3530.012 B92 0.346 0.008 B93 0.525 0.004 B94 0.416 0.005 B95 0.438 0.009B96 >9.1 0.118 B97 0.476 0.0215 B98 0.910 0.013 B99 0.302 0.057 B1000.567 0.014 B101 0.471 0.015 B102 0.209 0.01025 B103 0.241 0.046 B1041.374 0.162 B105 0.762 0.02 B106 0.247 0.017 B107 0.239 0.004 B108 0.094<0.003 B109 0.118 <0.003 B110 0.148 0.003 B111 0.685 0.06 B112 0.380.026 B113 2.73 0.109 B114 1.440 0.071 B115 0.468 0.023 B116 0.304 0.022B117 1.212 0.008 B118 0.498 0.01 B119 0.124 0.006 B120 0.088 0.006 B1213.357 0.075 B122 0.425 0.024 C1 0.300 0.0190 C2 1.218 0.243 C3 3.3771.207 C4 3.001 0.028 C5 5.510 0.095 C6 4.259 0.181 C7 >9.1 C8 0.3740.013 C9 0.185 0.006 C10 0.147 0.015 C11 0.353 0.023 C12 0.303 0.116 C130.667 0.077 C14 0.250 0.005 C15 0.383 0.017 C16 0.478 0.015 C17 0.3580.008 C18 0.425 0.012 C19 0.488 0.021 C20 0.193 0.006 C21 0.698 0.028C22 0.202 0.02 C23 0.658 0.032 C24 0.560 0.003 C25 0.474 0.0125 C266.946 3.219 D1 0.424 0.005 D2 2.743 0.149 D3 4.418 0.219 D4 0.236 D50.802 0.042 D6 0.650 0.009 D7 0.234 D8 0.463 0.011 D9 0.494 0.015 D100.385 0.062 D11 0.442 0.019 D12 0.142 <0.003 D13 0.5225 0.005 D14 0.415D15 0.711 0.058 D16 0.503 0.007 D17 0.705 0.012 D18 0.281 0.019 D190.273 0.013 D20 0.206 0.007 D21 0.519 0.041 D22 1.018 0.024 D23 2.5340.214 D24 >9.1 7.085 D25 >9.1 2.693 D26 1.236 0.028 D27 1.574 0.064 D280.411 0.005 D29 0.534 0.003 D30 5.150 0.127 D31 0.135 0.009 D32 0.0940.009 D33 0.117 0.008 D34 0.273 0.009 D35 0.414 <0.003 D36 0.346 D370.298 0.127 D38 0.516 0.018 D39 0.987 0.036 D40 0.7045 0.025 E1 0.0980.004 E2 E3 0.494 <0.003 E4 0.870 0.010 E5 E6 0.341 0.009 E7 0.133 0.022E8 E9 0.593 0.007 E10 0.229 0.013 E11 0.377 <0.003 F1 0.109 0.003 F20.725 0.005 F3 0.337 0.006 F4 0.577 0.003 F5 0.444 0.025 F6 0.783 0.003F7 0.071 <0.003 F8 0.038 0.005 F9 0.028 <0.003 F10 0.039 <0.003 F110.078 0.076 F12 0.440 <0.003 F13 0.704 0.024 G1 0.049 0.017 G2 1.1950.207 G3 0.372 0.056 H1 0.850 0.008 H2 0.913 0.013 H3 0.559 0.008 H40.8475 0.011 H5 0.477 0.011 H6 0.846 0.053 H7 0.703 0.005 H8 0.521 0.007H9 0.616 0.011 H10 0.727 0.018 H11 0.299 0.008 H12 0.530 0.007 H13 0.3870.009 H14 0.103 0.005 H15 0.724 0.008 H16 0.871 0.016 I1 0.842 0.018 I20.308 0.005 I3 0.836 0.006 J 0.998 0.065 K 0.393 <0.003 L 0.538 0.013 M0.304 0.006 N 0.838 0.012 O1 4.574 0.053 O2 5.692 0.049 P 0.954 0.072 Q1.144 0.014 R 0.602 0.011 S 0.612 0.036 T 1.876 0.076 U 0.192 0.015 V3.196 0.079 W 2.979 0.056 X 3.575 0.061 Y 1.925

Cellular Assays

Cell PI3Kδ/ RWB/IC50 Example IC50 [umol l−1] CD86 [nmol l−1] A1 0.043 37B1 0.154 29 C1 0.081 68 D1 0.147 84 E1 0.007 78 E3 0.018 14 F1 0.011 7F3 0.050 F7 0.018 40 Q 0.145 37

The invention claimed is:
 1. A method of inhibiting the activity ofPI3Kδ isoform in a subject, wherein the method comprises administeringto the subject a therapeutically effective amount of a compoundaccording Formula (I):

or a pharmaceutically acceptable salt thereof, wherein Y is selectedfrom O or NH; V is selected from CR⁵ or N; W is selected from CH₂, or O;U is selected from N or CH; Q is selected from N or CR₆; wherein U and Qare not both N; R¹ is selected from phenyl, pyridyl, pyrimininyl,pyrazinyl, pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl,1,3,5-triazinyl, or —X—R⁴ wherein X is selected from C(O), S(O)₂ or CH₂and R⁴ is selected from C₁-C₈-alkyl, halo-C₁-C₈-alkyl,hydroxy-C₁-C₈-alkyl, C₁-C₈-alkoxy-C₁-C₈-alkyl, cyano-C₁-C₈-alkyl,N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl, C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl,phenyl, heterocyclyl, heterocyclyl-oxy, heterocyclyl-C₁-C₈-alkyl,C₃-C₁₂-cycloalkyl, C₃-C₁₂-cycloalkyl-oxy, C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl,heteroaryl, heteroaryl-oxy, heteroaryl-C₁-C₈-alkyl, hydroxy,C₁-C₈-alkoxy, amino, N—C₁-C₈-alkyl-amino or N,N-di-C₁-C₈-alkyl-amino,wherein C₁-C₈-alkyl in N—C₁-C₈-alkyl-amino and inN,N-di-C₁-C₈-alkyl-amino may be unsubstituted or substituted by halogen,hydroxy or C₁-C₄-alkoxy, wherein C₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyland in C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or substitutedby 1-5 substituents selected from halogen, hydroxy or C₁-C₄-alkoxy;wherein ‘heterocyclyl’ is a 3 to 7 membered saturated or partiallyunsaturated monocyclic ring system containing 1 to 3 heteroatomsselected from N, O or S, each of which is unsubstituted or substitutedby 1-5 substituents selected from oxo, halogen, C₁-C₈-alkyl,halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl, C₁-C₈-alkoxy,C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’ can beattached at a heteroatom or a carbon atom and where the N and/or Sheteroatoms can also optionally be oxidized to various oxidation states,wherein ‘heteroaryl’ is a 3 to 7 membered fully unsaturated monocyclicring system containing 1 to 3 heteroatoms selected from N, O or S, orpyrazolo[1,5-a]pyrimidine or imidazo[2,1-b]thiazole, each of which isunsubstituted or substituted by 1-5 substituents selected from halogen,C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heteroaryl’ can be attachedat a heteroatom or a carbon atom and where the N and/or S heteroatomscan also optionally be oxidized to various oxidation states; R⁶ isselected from hydrogen, halogen, C₁-C₄-alkyl, halo-C₁-C₄-alkyl,C₁-C₄-alkoxy, C₁-C₄-alkyl-sulfonyl, C₁-C₄-alkyl-sulfinyl,C₁-C₄-alkyl-sulfanyl, halo-C₁-C₄-alkoxy, C₁-C₄-alkoxy-C₁-C₄-alkyl,amino, N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino; R⁷ is selectedfrom hydrogen, halogen, cyano, nitro, C₁-C₄-alkyl, halo-C₁-C₄-alkyl,C₁-C₄-alkoxy, N(R⁸)₂-sulfonyl, C₁-C₄-alkyl-sulfonyl,C₁-C₄-alkyl-sulfonyl-amino, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,N—C₁-C₈-alkyl-amino, or N,N-di-C₁-C₈-alkyl-amino; or R⁶ and R⁷, togetherare CH═CH—CH═CH, wherein R⁸ is independently selected from hydrogen,C₁-C₄-alkyl, C₁-C₄-alkoxy or two R⁸ together with the nitrogen they areattached to form a 4 to 7 membered heterocyclic ring containing 1-2heteroatoms selected from N, O, S, which is unsubstituted or substitutedby 1-3 substituents selected from C₁-C₄-alkyl; R⁵ is independentlyselected from H, D, F or C₁-C₂-alkyl; R³⁰ is independently selected fromH, D or F.
 2. A method of ameliorating a disorder or a disease selectedfrom rheumatoid arthritis (RA), systemic lupus erythematosus (SLE),multiple sclerosis (MS), myasthenia gravis (MG), Sjögren's syndrome (SS)transplant rejection, lymphoctic leukemia, non-Hodgkin lymphoma, andlymphomas comprising administering to a subject suffering from thedisease or disorder a therapeutically effective amount of a compoundaccording Formula (I):

or a pharmaceutically acceptable salt thereof, wherein Y is selectedfrom O or NH; V is selected from CR⁵ or N; W is selected from CH₂, or O;U is selected from N or CH; Q is selected from N or CR₆; wherein U and Qare not both N; R¹ is selected from phenyl, pyridyl, pyrimininyl,pyrazinyl, pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl,1,3,5-triazinyl, or —X—R⁴ wherein X is selected from C(O), S(O)₂ or CH₂and R⁴ is selected from C₁-C₈-alkyl, halo-C₁-C₈-alkyl,hydroxy-C₁-C₈-alkyl, C₁-C₈-alkoxy-C₁-C₈-alkyl, cyano-C₁-C₈-alkyl,N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl, C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl,phenyl, heterocyclyl, heterocyclyl-oxy, heterocyclyl-C₁-C₈-alkyl,C₃-C₁₂-cycloalkyl, C₃-C₁₂-cycloalkyl-oxy, C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl,heteroaryl, heteroaryl-oxy, heteroaryl-C₁-C₈-alkyl, hydroxy,C₁-C₈-alkoxy, amino, N—C₁-C₈-alkyl-amino or N,N-di-C₁-C₈-alkyl-amino,wherein C₁-C₈-alkyl in N—C₁-C₈-alkyl-amino and inN,N-di-C₁-C₈-alkyl-amino may be unsubstituted or substituted by halogen,hydroxy or C₁-C₄-alkoxy, wherein C₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyland in C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or substitutedby 1-5 substituents selected from halogen, hydroxy or C₁-C₄-alkoxy;wherein ‘heterocyclyl’ is a 3 to 7 membered saturated or partiallyunsaturated monocyclic ring system containing 1 to 3 heteroatomsselected from N, O or S, each of which is unsubstituted or substitutedby 1-5 substituents selected from oxo, halogen, C₁-C₈-alkyl,halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl, C₁-C₈-alkoxy,C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’ can beattached at a heteroatom or a carbon atom and where the N and/or Sheteroatoms can also optionally be oxidized to various oxidation states,wherein ‘heteroaryl’ is a 3 to 7 membered fully unsaturated monocyclicring system containing 1 to 3 heteroatoms selected from N, O or S, orpyrazolo[1,5-a]pyrimidine or imidazo[2,1-b]thiazole, each of which isunsubstituted or substituted by 1-5 substituents selected from halogen,C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heteroaryl’ can be attachedat a heteroatom or a carbon atom and where the N and/or S heteroatomscan also optionally be oxidized to various oxidation states; R⁶ isselected from hydrogen, halogen, C₁-C₄-alkyl, halo-C₁-C₄-alkyl,C₁-C₄-alkoxy, C₁-C₄-alkyl-sulfonyl, C₁-C₄-alkyl-sulfinyl,C₁-C₄-alkyl-sulfanyl, halo-C₁-C₄-alkoxy, C₁-C₄-alkoxy-C₁-C₄-alkyl,amino, N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino; R⁷ is selectedfrom hydrogen, halogen, cyano, nitro, C₁-C₄-alkyl, halo-C₁-C₄-alkyl,C₁-C₄-alkoxy, N(R⁸)₂-sulfonyl, C₁-C₄-alkyl-sulfonyl,C₁-C₄-alkyl-sulfonyl-amino, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,N—C₁-C₈-alkyl-amino, or N,N-di-C₁-C₈-alkyl-amino; or R⁶ and R⁷, togetherare CH═CH—CH═CH, wherein R⁸ is independently selected from hydrogen,C₁-C₄-alkyl, C₁-C₄-alkoxy or two R⁸ together with the nitrogen they areattached to form a 4 to 7 membered heterocyclic ring containing 1-2heteroatoms selected from N, O, S, which is unsubstituted or substitutedby 1-3 substituents selected from C₁-C₄-alkyl; R⁵ is independentlyselected from H, D, F or C₁-C₂-alkyl; R³⁰ is independently selected fromH, D or F.
 3. The method of claim 2, wherein the compound of Formula Iis a compound of formula (Ic′):


4. The method of claim 2, wherein the compound of Formula I is acompound of formula (Id′)


5. The method of claim 2, wherein the compound of Formula I is acompound of formula (Ie′)


6. The method of claim 2, wherein the compound of Formula I is acompound of formula (If′)


7. The method of claim 4, wherein the compound of Formula I is acompound of formula (If′)


8. The method of claim 2, wherein the compound of Formula I is acompound in which R¹ is selected from phenyl, pyridyl, pyrimininyl,pyrazinyl, pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl,1,3,5-triazinyl, or —X—R⁴, wherein R⁴ is selected from C₁-C₈-alkyl,hydroxy-C₁-C₈-alkyl, C₁-C₈-alkoxy-C₁-C₈-alkyl, cyano-C₁-C₈-alkyl,N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl, C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl,phenyl, heterocyclyl, heterocyclyl-C₁-C₈-alkyl, C₃-C₁₂-cycloalkyl,heteroaryl, heteroaryl-C₁-C₈-alkyl, C₁-C₈-alkoxy, wherein C₁-C₈-alkyl inN—C₁-C₈-alkyl-amino and in N,N-di-C₁-C₈-alkyl-amino may be unsubstitutedor substituted by halogen, hydroxy or C₁-C₄-alkoxy, whereinC₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyl and inC₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or substituted byhalogen, hydroxy or C₁-C₄-alkoxy; wherein ‘heterocyclyl’ is a 3 to 7membered saturated or partially unsaturated monocyclic ring systemcontaining 1 to 3 heteroatoms selected from N, O or S, which isunsubstituted or substituted by 1-5 substituents selected from oxo,halogen, C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’ can beattached at a heteroatom or a carbon atom and where the N and/or Sheteroatoms can also optionally be oxidized to various oxidation states,wherein ‘heteroaryl’ is a 3 to 7 membered fully unsaturated monocyclicring system containing 1 to 3 heteroatoms selected from N, O or S, orpyrazolo[1,5-a]pyrimidine or imidazo[2,1-b]thiazole, each of which isunsubstituted or substituted by 1-5 substituents selected from halogen,C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heteroaryl’ can be attachedat a heteroatom or a carbon atom and where the N and/or S heteroatomscan also optionally be oxidized to various oxidation states.
 9. Themethod of claim 2, wherein the compound of Formula I is a compound inwhich R¹ is selected from —X—R⁴, wherein R⁴ is selected fromC₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, C₁-C₈-alkoxy-C₁-C₈-alkyl,cyano-C₁-C₈-alkyl, N,N-di-C₁-C₄-alkyl-amino-C₁-C₈-alkyl,C₁-C₄-alkyl-sulfonyl-C₁-C₈-alkyl, phenyl, heterocyclyl,heterocyclyl-C₁-C₈-alkyl, C₃-C₁₂-cycloalkyl, heteroaryl,heteroaryl-C₁-C₈-alkyl, C₁-C₈-alkoxy, wherein C₁-C₈-alkyl inN—C₁-C₈-alkyl-amino and in N,N-di-C₁-C₈-alkyl-amino may be unsubstitutedor substituted by halogen, hydroxy or C₁-C₄-alkoxy, whereinC₃-C₁₂-cycloalkyl in C₃-C₁₂-cycloalkyl and inC₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or substituted byhalogen, hydroxy or C₁-C₄-alkoxy, wherein ‘heterocyclyl’ is a 3 to 7membered saturated or partially unsaturated monocyclic ring systemcontaining 1 to 3 heteroatoms selected from N, O or S, which isunsubstituted or substituted by 1-5 substituents selected from oxo,halogen, C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heterocyclyl’ can beattached at a heteroatom or a carbon atom and where the N and/or Sheteroatoms can also optionally be oxidized to various oxidation states,wherein ‘heteroaryl’ is a 3 to 7 membered fully unsaturated monocyclicring system containing 1 to 3 heteroatoms selected from N, O or S, orpyrazolo[1,5-a]pyrimidine or imidazo[2,1-b]thiazole, each of which isunsubstituted or substituted by 1-5 substituents selected from halogen,C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein ‘heteroaryl’ can be attachedat a heteroatom or a carbon atom and where the N and/or S heteroatomscan also optionally be oxidized to various oxidation states; R⁶ isselected from halogen, C₁-C₄-alkoxy, C₁-C₄-alkyl-sulfonyl orhalo-C₁-C₄-alkoxy and R⁷ is selected from hydrogen, halogen, cyano,C₁-C₄-alkyl, halo-C₁-C₄-alkyl or C₁-C₄-alkoxy.
 10. The method of claim2, wherein the compound of Formula I is a compound in crystalline form.